Fig. 3: INH^R-katG utilizes alternative redox detoxification pathways to compensate for the loss of katG.

Susceptibility of M. tuberculosis DS-parent and INH^R-katG to growth inhibition by (a) H₂O₂ and (b) ascorbic acid (mean ± extrema of two biological replicates, n = 3 independent experiments). c CellRox based detection of reactive oxygen stress in M. tuberculosis DS-parent and INH^R-katG when exposed to H₂O₂ (mean ± SD of three biological replicates, n = 2 independent experiments). d Changes in gene expression INH^R-katG relative to the DS-parent and increased depletion of gRNA abundance in INH^R-katG relative to DS-parent. Genes are named using gene name or rv number. gRNAs that are more depleted in INH^R-katG are presented as white dots, with the positioning of each dot denoting the level of increased depletion on a log2FC scale. Only gRNAs that show a statistically significant depletion on day 14+ ATc-300 are presented. e–h INH^R-katG is more sensitive to iron dysregulation: (e) Abundance of gRNAs targeting ideR throughout the WG-CRISPRi screen as detected by amplicon sequencing. Data is presented for cultures exposed to ATc-300. f Growth of M. tuberculosis DS-parent and INH^R-katG expressing gRNA targeting ideR (mean ± SD of two biological replicates, n ≥ 3). The (gx) after each gRNA denotes the specific gRNA targeting ideR. g Intracellular survival of M. tuberculosis DS-parent and INH^R-katG expressing gRNA targeting ideR (mean ± SD of three biological replicates, n = 2 independent experiments). h Susceptibility of M. tuberculosis DS-parent and INH^R-katG to 2.5 mM FeCl as determined by CFU/ml (mean ± SD of three biological replicates, n = 2 independent experiments). i Growth kinetics of M. tuberculosis DS-parent and INH^R-katG expressing gRNA targeting ideR, Bacterial growth was measured by OD600. All strains were grown in 7H9 media with ATc-300 and back diluted 1/20 with fresh media on days 5 and 10. Data is mean ± SD of three biological replicates, n = 2 independent experiments. j The ability of M. tuberculosis DS-parent and INH^R-katG to deplete oxygen during the transition to hypoxia was detected using PreSens oxygen sensing spots. Data represents the individual oxygen consumption curves of two biological replicates from a representative experiment. k The viability of M. tuberculosis DS-parent and INH^R-katG once the concentration of dissolved oxygen was less than 1% was detected by plating for viable colonies. Data is shown as the mean ± SD of three biological replicates, n = 2 independent experiments. Source data are provided as a Source Data file.