Fig. 4.
C10 inhibits respiration in Mtb. (A) RNA-seq was performed on Mtb treated with 25 μM C10 for 48 h in aerobic conditions. The functional categories based on gene annotations in Mycobrowser for the genes significantly (Padj < 0.05) up-regulated by >1.5-fold are presented in a pie chart. (B) Mtb was pretreated for 4 h with DMSO, 2.5 μg/mL INH, 1 μg/mL CFZ, or 50 μM C10, followed by the addition of methylene blue for an additional 16 h. n = 3. (C) The viable bacteria from B were enumerated. (D) Mtb was treated with increasing concentrations of C10 and monitored for respiration in the MABA, and the IC50 ± SD was calculated with GraphPad Prism. n = 3. (E and F) WT Mtb was incubated with 5 or 25 μM C10 ± 0.25 μg/mL INH for 24 h, and either ATP levels were measured in relative luminescence units (RLU) using BacTiter Glo (E), or ROS levels were quantified by CellROX Green fluorescence in arbitrary units (AU) (F). In F, 1 μg/mL CFZ was included as a positive control. In C–F, values are mean ± SEM. *P < 0.05; ****P < 0.0001; ns, not significant by one-way ANOVA with Tukey’s test. Relevant comparisons are indicated. Complete statistics are provided in SI Appendix, Table S1.