Figure 5.
MV induction in other mycolic acid-containing bacteria
(A–T) The panels (A–G, H–N, and O–T) correspond to Mycobacterium smegmatis MC2155, R. erythropolis PR4, and Rhodococcus equi IFO3730, respectively. (A, H, and O) M. smegmatis, R. erythropolis, and R. equi were cultured under various conditions and then their MV release were measured. M. smegmatis and R. erythropolis were cultured in synthetic minimum media, whereas R. equi was cultured in LB medium, with and without biotin supplementation, due to extremely low growth in synthetic minimum medium (details of growth conditions are described under Methods). All values indicated by the bars represent the mean value ± SD for three experiments. p values were calculated using unpaired t test with Welch's correction. DCW, dried cell weights. (B, I, and P) TEM images of MVs of the mycolic acid containing bacteria are shown. Scale bars, 200 nm. (C–F, J–M, and Q–S) Particle size distributions of the above MVs. Black lines indicate the mean values of the concentrations of the detected particles in MV solutions. Red regions indicate SD of the mean values. (G, N, and T) Thin-layer chromatography profiles of the above MVs. Lipids were processed using chloroform-methanol-water (65:25:4, v/v). N, no treatment condition; B, biotin-deficient condition in M. smegmatis and R. erythropolis, or without biotin supplemented in R. equi; M, MMC treatment condition; P, penicillin G treatment condition. Black star indicates apolar lipids including mycolic acid esters. Black circle indicates polar lipids including phospholipids.