Fig. 1: Genome-wide CRISPR screen to identify host determinants of tuberculosis restriction in macrophages.
A Mycobacterium tuberculosis (Mtb) induced cytotoxicity in Hoxb8 bone marrow-derived macrophages (BMDMhs) at the indicated multiplicity of infections (MOI) 4 days post infection. Cytotoxicity was quantified by measuring the activity of released lactate dehydrogenase (LDH) in cell supernatants as compared to 100% LDH activity in detergent-lysed cells; n = 4 biological replicates. **P < 0.01; ***P < 0.001, one-way ANOVA alongside Dunnett’s multiple comparison test. Data are presented as mean values ± SD. B A further quantification of Mtb induced cytotoxicity in macrophages at indicated MOIs by plating colony forming units (CFUs) in cell supernatants from experimental conditions as in (A); n = 5 biological replicates. **P < 0.01; ***P < 0.001, one-way ANOVA alongside Dunnett’s multiple comparison test. Data are presented as mean values ± SD. C Schematic workflow for performing CRISPR screen to identify host determinants of Mtb restriction in macrophages. Part of this graphic was created in BioRender. Lee, B. (2024) BioRender.com/t32i914. D Volcano plot showing hits from the screen. For each gene (represented by dots), enrichment or depletion is shown in the x axis as log2 fold change while the y axis shows the corresponding −log10 p value. Significantly enriched and depleted hits are shown in red and blue colors, respectively, while unchanged hits are in gray. For significant hits, an adjusted p value of <0.05 was used as a cutoff. Screens were carried out in three independent replicates. Source data are provided as a Source Data file.
