Extended Data Fig. 4: Specific Targeting of the H526Y phenotypic class of RifR mutants.
Increased sensitivity of RifR cells harbouring alleles H526Y, D516G, and S522F to A22 (1 μg/ml), 5FUd (5 μg/ml) and high temperature. The S531F is included as an example of a RifR allele that does not share the same phenotypic profile. Mean and standard errors of all plating efficiency data are based on colony forming unit values (c.f.u) from three biological triplicates. * p-value <0.05, ** p-value <0.01, *** p-value <0.001. p-values are based on the student t-test (two-sided, equal variance) b, Growth of wild type and rpoB H526Y cells in LB supplemented with increasing concentration of human serum, with or without subinhibitory concentration of A22. c, Bacterial load in the peritoneal lavage, spleen, and liver in mice 24 h after infection with either wild type (triangle-shaped symbols) or H526Y cells (circle-shaped symbols). For each strain, the two groups of mice (n = 10 mice/group) received either no drug (clear symbols) or 20 mg/kg A22 one-hour postinfection (filled symbols). The median for each group is indicated with a solid horizontal line. Dotted lines indicate limit of detection. * p-value <0.05, ** p-value <0.01, n.s. not significant. d, Phenotypic profiling of wild type and rpoB H526Y cells against a collection of 56 antibiotics and antimicrobials with a known mode of action. colour code is according to the cellular target of each drug. Abbreviations of each class of drugs: Aminoglycosides (AG), Cell wall synthesis and division inhibitors (CW), Quinolones (QUI), genotoxic compounds (GNTX), DNA polymerase starving molecules (REP), Tetracyclines (TETC), Macrolides (MCLD), rifamycins (RIF), Antimicrobial peptides (AMP), and in a miscellaneous group (MICEL). Arrows point to antibiotics that were selected for MIC analysis across the nine different strains that are presented in Fig.3i.
