Direct Visualization by Cryo-EM of the Mycobacterial Capsular Layer: A Labile Structure Containing ESX-1-Secreted Proteins (original) (raw)
Figure 3
Effect of detergent on the localization of capsular components and the detection of ESX-1 proteins.
M. smegmatis (A and B), M. tuberculosis (D and E) and M. marinum (G and H) were grown under both perturbed (A,D and G) and unperturbed conditions (B,E and H), fixed and probed with anti-α-glucan (A–B) and anti PIM/capLAM (D–E) to demonstrate that this extra layer is of capsular origin. (G–H) demonstrates the presence of anti-EspE on bacterial surface/capsule. Histogram (C,F and I) respectively displays the variations in anti glucan, anti PIM/capLAM and anti-EspE labeling of bacteria due to change in the culturing conditions using M. smegmatis ΔPimE k.o. and M. marinum fas 3.1 eccCb1 mutant as controls. The ordinate (defined as percentage of bacteria with ≥10 gold particles) represents the average ± standard error of labeled cells from 3 independent experiments. Scale bars: 250 nm.