DG167 inhibits mycolic acid biosynthesis in vivo. (C and D) Normal-phase TLC analysis of MAMEs and FAMEs from wild-type M. tuberculosis (H37Rv) and DG167-resistant M. tuberculosis (isolate DRM167-32x2; Table 1). The cultures were treated with increasing concentrations of DG167 and inactive analog 5a-2. Total lipids were extracted, subjected to methyl esterification after [14C]acetate incorporation, and resolved by TLCs. RFU, relative fluorescence units. (A) M. tuberculosis H37Rv. (B) M. tuberculosis DRM167-32x2 isolate. (C) Densitometric analysis of MAMEs and FAMEs from panel A using ImageQuant (GE Healthcare). (D) Densitometric analysis of MAMEs and FAMEs from panel B. Equal counts (20,000 cpm) were loaded, and the TLC was developed using hexane/ethyl acetate (19:1 [vol/vol], 2 runs).