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Roy, A.

Publications and source records attributed to Roy, A..

At least 37 records · Page 2Linked to original sources

Self-supervised learning for a gene program-centric view of cell states

Single-cell omics has extended the biological interrogation of cell state from examining the expression of individual genes to unbiased profiling of tens of thousands of genes at once. However, extracting biological insights from such high-dimensional data remains challenging. To enable downstream analyses, many computational approaches compress cell state into a single latent representation. This can obscure the structure of underlying gene programs (GP), defined as coordinated sets of biologically related genes, such as signalling pathway response modules or transcription factor targets. Here, we present Tripso, a self-supervised transformer deep learning model which learns multiple GP-specific embeddings from predefined GPs, while also enabling the discovery of novel, data-driven GPs. Tripso facilitates principled comparisons across development, disease, and experimental systems. Firstly, in a dataset of human hematopoietic cells spanning prenatal development through adulthood and aging and including newly generated data, Tripso resolved age-specific GP patterns, including elevated JAK-STAT activity in pediatric hematopoietic cells and postnatal shifts in IKZF1 GP activity during B cell differentiation. Secondly, leveraging Tripso GP embeddings and comparing in vivo to in vitro data, we hypothesized and experimentally validated that inhibition of the SEC61 translocon improved maintenance of hematopoietic stem cells in culture. Finally, Tripsos capacity for data-driven GP discovery revealed a previously uncharacterized tissue-resident memory T cell GP with increased activity in atopic dermatitis. Its spatial co-localization with sebaceous gland-associated immune niches was demonstrated in spatial transcriptomic and proteomic data. Thus, by moving beyond single embeddings of cellular states, Tripso enables interpretable and actionable discoveries, demonstrating how GP-centric modelling can generate hypotheses with substantial biomedical relevance. By anchoring cellular representations in meaningful GPs, Tripso establishes a principled and biologically grounded framework towards the development of interpretable virtual cell models.

bioinformatics↗

Comprehensive mapping of Neurofibromin 1 (NF1) expression in developing mouse brain

Neurofibromin (NF1) is a critical negative regulator of the RAS-RAF-ERK pathway, mutations in which have been clinically implicated in various neurodevelopmental disorders. However, the lack of a high-resolution spatiotemporal map has obscured the understanding of why specific cell populations and developmental processes are uniquely vulnerable to NF1 loss. In this study, we present a comprehensive atlas of NF1 expression in the developing mouse brain. Using in situ hybridization and immunohistochemistry, we characterized NF1 distribution from early embryonic stages through postnatal maturation. We further integrated these findings with single-nuclei RNA-sequencing (snRNA-seq) datasets from adult mouse brain to achieve higher resolution. Our results reveal a previously undocumented graded expression pattern of NF1 across various brain regions and cell lineages. This comprehensive study will not only help understand the fundamental role of NF1 in brain development but will also be pivotal in providing a framework for studying NF1-associated brain disorders. Summary statementGraded expression of Neurofibromin (NF1) identified across embryonic and postnatal stages reveals unique sub-lineages of neural cells in different parts of the developing mouse brain.

neuroscience↗

Repeated trends in altitudinal gradients of diversity: how habitat filtering and biotic interactions structure ecological communities

Understanding how biodiversity is structured along tropical elevational gradients requires disentangling the relative roles of regional evolutionary history and local processes shaping ecological assemblies. Here, Ithomiini butterfly communities were studied along repeated elevational gradients in two Neotropical regions with contrasting evolutionary histories: the Amazonian Andes and the Guiana Shield. The study tested whether similar elevational patterns of taxonomic, mimetic, and phylogenetic structure emerge despite distinct regional species pools, and whether abiotic and biotic factors contribute to shaping these patterns. Despite marked regional differences in overall richness, consistent elevational patterns emerged across both regions. Taxonomic and mimetic richness increased with elevation and were accompanied by stronger phylogenetic clustering, indicating that similar habitat filtering processes operate along altitudinal gradients irrespective of regional context. Phylogenetic {beta}-diversity was predominantly driven by lineage turnover, particularly in the Andes, highlighting the role of elevational gradients in promoting replacement of phylogenetically distinct lineages rather than simple species loss. These shared patterns suggest that altitude has a strong and repeatable effect on community structure, with habitat filtering acting locally on regionally distinct species pool. Abiotic factors such as temperature appeared to constrain species distributions at broad spatial scales, whereas biotic interactions acted more locally. In particular, butterfly diversity was positively associated with potential host plant richness and predation pressure, indicating that ecological interactions can further shape local community composition once broad-scale environmental constraints are accounted for. By integrating phylogenetic structure, biotic interactions, and environmental gradients across regions with contrasting evolutionary histories, this study shows how regional species pools and local ecological filtering jointly shape tropical biodiversity and highlights that similar elevational assembly processes could arise independently across the Neotropics.

ecology↗

Epigenetic Silencing of Carotid Body TRPM7 Attenuates Hypertension in Obese Mice

Obesity is the most common cause of hypertension. We have previously shown that high levels of circulating leptin in diet-induced obese (DIO) mice induced hypertension by increasing expression of Transient Receptor Potential Melastatin-subfamily member 7 (TRPM7) in the carotid bodies (CB). In addition, we demonstrated in rat PC12 cells that leptin increases Trpm7 gene expression by inducing CpG site-specific demethylation within the 5 regulatory region containing a signal transducer and activator of transcription 3 (STAT3) binding site. This leptin-induced Trpm7 upregulation was prevented by inhibition of JAK-STAT3 signaling. Based on these findings, we hypothesized that reversing region-specific methylation at the Trpm7 promoter in the CB could attenuate obesity-associated hypertension. Compared with lean controls, DIO mice exhibited increased Trpm7 expression and the STAT3- binding site-specific promoter demethylation in the CB. Administration of methylated DNA oligonucleotides targeting the STAT3 binding site attenuated CpG site-specific DNA demethylation and reduced Trpm7 transcription in the CB of DIO mice. This intervention resulted in decreased carotid sinus nerve activity and reduced arterial blood pressure, especially during the light phase. Our results suggest that targeted modulation of CpG site-specific DNA methylation at the Trpm7 promoter using DNA oligonucleotide may represent a novel therapeutic strategy for obesity-induced hypertension.

physiology↗

Decoding Phonetic Features: Somatotopic and Sensorimotor Representations in Native and Non-native Consonant Perception

Speech perception relies on the integration of auditory and articulatory information, yet the precise role of motor regions remains debated. We combined behavioral measures and fMRI with multivariate pattern analyses to investigate cortical representations of native French and non-native Mandarin consonant perception under clear and noisy conditions. Cross-modal classification analysis showed that articulatory features of perceived degraded native bilabial and dental consonants are mapped somatotopically in right lip and tongue motor areas, which are also engaged in consonant production. These representations may support phoneme categorization by compensating for degraded input. Representational similarity analysis further revealed that a network encompassing bilateral temporal and frontal motor regions encodes phonetic features of native and non-native consonants, including place and manner of articulation. Our findings highlight that speech perception relies on embodied sensorimotor representations, which contribute to decoding phonetic features both within and across languages.

neuroscience↗

Predicting how perturbations reshape cellular trajectories with PerturbGen

A major challenge in biology is predicting how cells transition between states over time and how perturbations disrupt these transitions. Understanding such dynamics is critical for identifying interventions that reverse pathological programs or reprogram cells toward desired states. Although recent computational approaches can predict single-cell perturbation responses in silico, they cannot predict responses across dynamic cell trajectories, for example how early perturbations reconfigure later cell states. To address this gap, we introduce PerturbGen, a generative foundation model trained on over 100 million single-cell transcriptomes that predicts perturbation responses along cellular trajectories. PerturbGen predicts how genetic perturbation at source state shapes downstream states, alters gene programs and trajectories across time, for example in differentiation or disease progression. We apply PerturbGen to three newly generated multi-condition human single-cell datasets spanning immune responses, hematopoiesis and skin development. In an in vivo immune challenge, PerturbGen predicts that knocking out an IL1B signal in myeloid cells attenuates later cytokine-interferon programs, with downstream changes consistent with a reversal of IL-1{beta} stimulation signature. In hematopoiesis, anchoring perturbation-induced programs to human genetics enables simulation of monogenic blood disorders, recapitulates established disease-associated biology whilst systematically revealing lineage-specific programs, including in lineages where this was not previously possible. In skin organoids, PerturbGen predicted that Wnt activation enhances stromal differentiation recapitulating the trajectory observed in human prenatal skin, findings that were functionally validated by experimentally activating Wnt signaling. Together, PerturbGen extends modeling of gene perturbations from static to dynamic cellular systems. We envision PerturbGen enabling the creation of in silico, trajectory-aware perturbation atlases and virtual cells across diverse biological scenarios, supporting optimization of disease models and prioritization of candidate molecular interventions for therapeutic discovery.

bioinformatics↗

TAK1 regulates skeletal muscle mass, hypertrophic signaling, and metabolic homeostasis in male and female mice

Skeletal muscle is the most abundant tissue in the human body and is essential for locomotion and the regulation of whole-body metabolism. The maintenance of skeletal muscle mass is essential for health, yet the molecular and signaling mechanisms that control skeletal muscle mass remain poorly understood. Transforming growth factor-{beta}-activated kinase 1 (TAK1) is a key signaling protein that regulates multiple intracellular pathways. Recent studies have demonstrated that TAK1 is a critical regulator of skeletal muscle mass. However, the mechanisms by which TAK1 regulates muscle mass and whether its role is sex dependent remain incompletely understood. In this study, we show that targeted inactivation of TAK1 induces muscle atrophy more rapidly in male than in female mice. Loss of TAK1 activity also abolished mechanical overload-induced phosphorylation of p70S6K and rpS6, and the induction of myofiber hypertrophy in both sexes. RNA-Seq analysis further revealed that TAK1 inactivation in skeletal muscle disrupts the gene expression of various molecules involved in catabolic processes, calcium signaling, muscle structure development, and aerobic respiration. Moreover, TAK1 inactivation impairs fatty acid oxidation and promotes lipid accumulation in skeletal muscle of adult mice in a sex-independent manner. Collectively, our findings demonstrate that TAK1 regulates skeletal muscle mass and growth by coordinating distinct intracellular pathways in both male and female mice.

physiology↗

EnzFinder: a sustainable alternative to chemical synthesis

Enzymes have emerged as an important alternative to traditional catalysts in chemical industries over the past few decades owing to their sustainable nature. The application of enzymes in chemical synthesis relies on their ability to catalyse promiscuous reactions. Promiscuous activity of enzymes is an abundant phenomenon in nature; approximately 37% of Escherichia coli K12 enzymes show promiscuous activity. This highlights the vast expanse of chemical reactions that can be made biochemically feasible through selection of the correct candidate enzymes. Here, we present EnzFinder, a promiscuous enzyme prediction tool that filters candidate enzymes based on similarity in chemical transformation patterns and subsequently ranks them using substrate-product similarity, enabling enzyme prioritization up to the fourth level of EC classification without requiring sequence information. On a blind benchmarking set of 2,309 biochemical reactions, the method achieves substantially higher prediction accuracy than existing rule-based and deep-learning approaches, with improvements exceeding 20% at the sub-subclass level and significantly higher coverage at the fourth level. Application to industrially relevant reactions demonstrates EnzFinders ability to identify alternative enzymes with higher substrate similarity and improved kinetic potential. Furthermore, integration of EnzFinder with in silico retrosynthesis tools enables effective prioritization of enzymatic steps within hybrid chemical-biological pathways. Together, these results establish EnzFinder as a practical and interpretable tool for accelerating enzyme discovery and promoting greener, enzyme-driven synthesis routes.

synthetic biology↗

Structural determinants of broadly neutralizing human antibodies binding to morphological dengue virus variants.

Dengue virus (DENV) is a global health threat, with four antigenically distinct serotypes impeding vaccine and therapeutic development. We describe here, structural features of patient-derived, heavy chain-driven E-dimer-recognizing (HEDR) antibodies that achieve potent, pan-serotype activity by binding a conserved quaternary epitope on DENVs envelope (E) glycoprotein dimer. Single-particle cryo-EM structures of three HEDR broadly neutralizing antibodies (bnAbs) complexed to DENV delineate epitope engagement via E-glycoproteins fusion loop region and conserved glycans. We also identify capsule-shaped DENV, with helical cryo-EM reconstructions of Fab-bound tubular DENV from different strains revealing that the E-glycoprotein arrangement is determined by virus strain, not bnAb identity. Our findings elucidate key binding features of HEDR bnAbs and signify the need for intervention strategies capable of neutralizing diverse DENV morphologies.

biophysics↗

Dual targeting a LIN28B:β-catenin axis in acute myeloid leukaemia

BackgroundWnt/{beta}-catenin signalling is dysregulated in acute myeloid leukaemia (AML), where it lacks effective targeting strategies. Previously, we discovered that {beta}-catenin interacts with several RNA-binding proteins (RBP), indicating post-transcriptional influence which is yet to be therapeutically interrogated in AML. MethodsCo-immunoprecipitation confirmed protein interactions, and TCF/LEF reporters were used to assess Wnt signalling output in leukaemia cells. Regulatory crosstalk was assessed using immunoblotting and RT-qPCR approaches following lentiviral transduction of myeloid cell lines. Targeting of {beta}-catenin and LIN28B was tested through combinations of genetic and pharmacological inhibition in AML cells. ResultsThe most frequent RBP-binding motif amongst {beta}-catenin-bound mRNAs was the GGAG motif targeted by oncofetal miRNA-regulating RBP; LIN28B. {beta}-Catenin:LIN28B interactions were detected in lymphoid and myeloid cell lines, plus primary human CD34 fetal-liver HSCs. LIN28B positively regulated Wnt signalling output through LEF1 regulation involving a post-transcriptional let7 miRNA mechanism. Further miRNA sequencing of {beta}-catenin- and LIN28B-depleted myeloid cells revealed potential cooperative and antagonistic function in miRNA regulation. Finally, dual-targeting both {beta}-catenin and LIN28B through either genetic and/or pharmacological means preferentially reduced AML cell viability. ConclusionThe {beta}-catenin:LIN28B axis could represent a novel synthetically lethal relationship in AML which could be exploited in rare subtypes where LIN28B expression becomes reactivated.

cancer biology↗

Pol γ possesses separate metal binding sites for polymerase and strand displacement functions

Accurate replication of mitochondrial genome (mtDNA) integrity, which is essential for cellular metabolism and energy supply, relies primarily on DNA polymerase gamma (Pol {gamma}), Twinkle helicase, and mitochondrial single-stranded DNA binding protein (mtSSB). Twinkle alone exhibits little helicase activity while reports indicate that Pol {gamma} displays from modest to limited unwinding activity. This led us to dissect Pol {gamma} strand displacement activity using structural, biochemical and in silico approaches. Here, we show that human Pol {gamma} carries out robust strand displacement synthesis at physiological concentrations of divalent metal ions which reveals that distinct metal-binding sites can independently regulate DNA synthesis and unwinding activities. We further showed that Pol {gamma} can displace RNA/DNA hybrid with comparable efficiency as DNA/DNA duplex, representing a key implication on RNA primer removal to preserve mtDNA integrity. Our cryo-electron microscopy structures of Pol {gamma} complexed with a template containing downstream dsDNA and an incoming nucleotide revealed the structural mechanism for the strand displacement activity. We identified four conformational states that represent successive stages of DNA unwinding, accompanied by coordinated rearrangement of the downstream DNA and Pol {gamma} elements that mediate strand displacement. This work establishes biochemical and structural mechanisms of Pol {gamma} strand displacement activity, providing fundamental insight into human mitochondrial DNA replication and integrity. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=185 SRC="FIGDIR/small/701366v1_ufig1.gif" ALT="Figure 1"> View larger version (39K): org.highwire.dtl.DTLVardef@1be1dborg.highwire.dtl.DTLVardef@8912c5org.highwire.dtl.DTLVardef@12f5e75org.highwire.dtl.DTLVardef@e25d2a_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

Menin maintains enhancer-promoter interactions in a leukemia-specific manner

Inhibition of the protein-protein interaction between Mixed Lineage Leukemia (MLL) and Menin is a promising therapy for both high-risk MLL-rearranged and NPM1-mutant (NPM1c) acute leukemias, yet the mechanistic basis of this dependency in distinct contexts remains unclear. By comparing the transcriptional responses of MLL::AF4 and NPM1c leukemia models to Menin inhibition, we find broad, acute transcriptional dysregulation in MLL::AF4 cells, but minor transcriptional consequences in NPM1c cells, despite similarities in Menin promoter occupancy. Using high-resolution Micro Capture-C, we discover that Menin drives enhancer activity and maintains enhancer-promoter contacts in MLL::AF4 cells but not in NPM1c cells. Crucially, Menin is also essential for patient-specific enhancer function in primary MLL-rearranged leukemia samples. Proteomic analysis further demonstrates that Menin associates with distinct transcriptional and elongation complexes in MLL::AF4 compared to NPM1c cells, supporting a context-dependent mechanism of action. Together, these findings establish that Menin is not a uniform transcriptional cofactor, but a context-dependent regulator of enhancer connectivity, and identifies enhancer-promoter architecture as a selective vulnerability in MLL-rearranged leukemia.

cancer biology↗

Nascent extracellular matrix converts biomaterial cues into cell fate decisions

Hydrogels serve as powerful models for investigating cell-extracellular matrix (ECM) interactions. While chemical modifications are routinely used to tune hydrogel properties, it remains unclear whether these modifications mediate cell fate. Previous work has shown that cells deposit newly synthesized (nascent) ECM at the cell-hydrogel interface. Here, we demonstrate that this nascent ECM interface regulates how cells interpret chemical modifications. Using hydrogels with varied chemical modifications, we isolated the effects of chemical modification on nascent ECM and cell fate. Nascent ECM deposition increased as a function of hydrogel modification and with distinct matrisome compositions. While low modification hydrogels promoted cell differentiation, high modifications increased cell proliferation. Perturbing cell-nascent ECM interactions reversed this cell fate. Our findings reveal that nascent ECM regulates cell fate by converting hydrogel cues into signals that control cell fate. This tri-directional interplay among hydrogel chemical modifications, nascent ECM, and cell fate reframes how we interpret cell-hydrogel interactions.

bioengineering↗

QTLIT: A user-friendly Shiny app for Quantitative Trait Loci (QTL) analysis

Identification of Quantitative Trait Loci from DNA sequence data is a multistep process, which is difficult for the plant breeders. To make the QTL identification breeders friendly, we developed QTLIT, a tool comprised of popular R packages and open sources software under a single umbrella for QTL analysis that enables intuitive exploration, multidimensional analyses, and visualization of results generated from any kind of DNA sequence data that deals with Single Nucleotide Polymorphism (SNP) variation. This tool requires raw or processed FASTQ files and proceed it for SNP identification to further perform QTL analysis on genomic regions of interest along with phenotyping data of various pedigree-based mapping populations. QTLIT generates a full range of plots such as VCF quality, linkage map, LOD score etc. It has broad range of applications and options for different parameters which the user can adjust as per their requirements. Moreover, it has two separate interfaces where one can utilize this tool to generate alignment files and/or variant calling for other downstream analysis. QTLIT is a fast and intuitive QTL analysis tool suitable for a wide range of users due to its architecture and its user-friendly graphical interface. QTLIT source code is available at https://github.com/TKM-Lab/QTLIT and the user interface version is available on http://14.139.229.202:8080/QTLIT.

genomics↗

MDCompress: better, faster compression of molecular dynamics simulation trajectories

MotivationMolecular dynamics (MD) simulations model the physical movements of atoms in biomolecular systems over time, providing atomic-resolution insight into conformational changes, binding events, and dynamic behaviors that cannot be captured by static structures alone. As such, MD simulations are playing an increasingly important role in understanding the functional roles and molecular interactions of proteins. However, trajectories from these simulations can be extremely large, often reaching tens of gigabytes for a single simulation of modest duration. This creates substantial challenges for storage and data transfer, motivating efficient compression strategies. Furthermore, many downstream analyses require extraction of only a subset of frames or specific atoms from the full trajectory, so an ideal compression format should support rapid random-access decompression of such samplings without requiring full file decompression. ResultsHere, we introduce MDCompress, a new trajectory compression format and accompanying software implementation that meets these goals. MDCompress produces compressed trajectory files that are 15-37% smaller than those generated by the widely-used XTC format, while achieving faster compression and decompression speeds through efficient multithreading. Availability and ImplementationThe MDCompress software and library are released under an open license (BSD-3) and may be downloaded at https://github.com/refresh-bio/mdcompress Contacttwheeler@arizona.edu, sebastian.deorowicz@polsl.pl Supplementary informationSupplementary data are available online.

biochemistry↗

Metagenomic characterisation of fungal communities associated with Scots pine bark beetles: mechanisms of selective antagonism and monoterpene tolerance

Bark beetle-associated fungi contribute to beetle nutrition, detoxification, and interactions with conifer hosts; however, their composition and function across development and environments remain poorly understood. We characterised fungal communities of two pine-feeding Ips bark beetles, I. sexdentatus and I. acuminatus, in larval, pupal, and adult stages, and in wild versus laboratory populations, using high-throughput ITS2 amplicon sequencing combined with qPCR and functional assays. Both beetles harboured a stable core mycobiome dominated by Kuraishia, Ogataea, Ophiostoma, Graphilbum and Cyberlindnera, while adults showed species-specific differences and wild beetles, especially I. acuminatus, exhibited greater diversity than laboratory populations. Beetles shared more taxa with unfed control wood than with gallery wood, indicating acquisition during feeding and concurrent restructuring of the wood mycobiome. Monoterpene bioassays on beetle-associated yeasts revealed that mixtures of -pinene, 3-carene and terpinolene suppressed growth more strongly than single compounds, suggesting synergistic inhibition. Yeasts selectively antagonised entomopathogenic fungi and expressed complementary cell-wall-lytic and digestive activities, consistent with combined roles in pathogen suppression and plant-polymer deconstruction. Our results show that Ips mycobiomes are conserved yet dynamic across life stages and environments, and emphasise the importance of multi-terpene and interaction assays for understanding bark beetle-fungus-conifer interactions. ImportanceOur study reveals that pine-feeding bark beetles co-evolved in close association with a stable core mycobiota that supports nutrient acquisition, detoxification, and chemical signalling, while additional fungal partners shift in response to beetle development, environment, and host context. By demonstrating higher mycobiome diversity in beetle larvae and wild populations, substantial overlap between beetle- and wood-associated fungi, differential sensitivity to monoterpene blends and selective antagonism of yeasts towards entomopathogenic fungi, our study unravels key ecological and mechanistic principles shaping beetle-fungus assemblages under conifers, paving the way for more in-depth functional investigations.

microbiology↗

Functional implications of compartment-specific homeostatic regulation and the feasibility of independent local feedback signals

Neurons regulate their average firing rate by adjusting their synaptic strength and intrinsic excitability. Intracellular calcium is implicated as an error signal for both types of homeostatic regulation. Past studies have focused on global calcium but two properties cannot be independently regulated by one error signal. Here, we computationally tested the implications of local vs global feedback and the feasibility of local (compartment-specific) error signals based on spatially segregated calcium changes. Simulations in a simple two-compartment model confirmed that a perturbation applied to one compartment induces local compensation only if feedback is compartment-specific. Simulations in a biophysically detailed multicompartment model with realistic calcium handling confirmed that dendritic and somatic calcium signals remain relatively segregated and can, therefore, encode separate error signals. Strong perturbations (as often tested experimentally) triggered widespread compensation because local compensation was overwhelmed. Non-local compensation also occurred when the spatial segregation of calcium signals was weakened. Our results demonstrate the plausibility of compartment-specific feedback using calcium-based error signals. Furthermore, whereas local homeostatic regulation nullifies local perturbations through compensation within the affected compartment, non-local regulation causes widespread compensatory changes that, while restoring the neurons overall input-output relationship, distorts the input-output relationship of individual compartments, with potentially important consequences.

neuroscience↗

Repurposing ethacridine as a potent MMPL3 Inhibitor for the treatment of tuberculosis

Mycobacterium tuberculosis (Mtb), the pathogen responsible for tuberculosis, remains a major global health threat, particularly with the rise of multidrug-resistant and extensively drug-resistant strains. This has renewed interest in repurposing existing drugs and exploring new cellular targets. The mycobacterial cell envelope is a key barrier to antibiotics and an attractive site for therapeutic intervention. In this study, we identify the FDA-approved drug ethacridine as a strong inhibitor of MmpL3, an essential transporter required for exporting trehalose monomycolate and building the cell wall. Computational docking and molecular dynamics indicate that ethacridine engages the MmpL3 binding pocket at residues also targeted by SQ109. Ethacridine shows potent activity against drug-sensitive and resistant Mtb isolates, with an MIC of 1 g/mL, and remains effective against non-replicating bacteria and intracellular infection. Ethacridine-resistant mutants, overexpression strains, and a spheroplast TMM-flipping assay confirm MmpL3 as the target. The compound also disrupts the membrane potential, and flow-cytometry assays

microbiology↗