Figure 3. Accumulation of TMMs in the MmpL3-depleted strain.

(A) Lipids were extracted from M. tuberculosis TB416, grown in the absence and presence of ATc (ATc−/+), with CHCl₃/CH₃OH (1:2–1x; 2:1–2x), then separated by TLC in the solvent CHCl₃/CH₃OH/H₂O (20:4:0.5) and detected with 10% (w/v) CuSO4 in 8% (v/v) H₃PO₄ for the complete lipid profiles. From the same cultures, delipidated cells, as well as lipid fractions were saponified with 15% TBAH. Free fatty/mycolic acids were derivatized to methylesters, analysed by TLC in n-hexane:ethyl acetate (95:5, 3x) and detected with a CuSO₄ detection system. (B) Mycolic acids bound to cell wall. (C) Fatty and mycolic acids from extractable lipids. Samples are in duplicates. H37Rv was used as a control. TDM: trehalose dimycolate; TMM: trehalose monomycolate; PE: phosphatidylethanolamine; CL: cardiolipin; FAME: fatty acid methyl ester; α, methoxy, keto: respectively alpha-, methoxy- and keto-mycolic acids (MAME, mycolic acid methyl ester).