Figure 3
The interaction of sortilin with GGAs/AP-1 and the retromer complex is required for phagosome maturation and mycobacterial restriction of growth.
(A) RAW 267.4 cells expressing the indicated proteins and infected with BCG-DsRed (white) for 2 h of uptake, followed by 1 h of chase were fixed and stained for LAMP-2 (red). Scale bar: 10 μm. (B) Quantitative analysis of fluorescence signal intensity for LAMP-2 association with BCG phagosomes. The numbers in the graph correspond to the mean of the fluorescence signal of LAMP-2 associated to BCG phagosome (a.u = arbitrary units) in the transfected cells. Data represents the Mean ± S.E.M of four independent experiments. At least 150 phagosomes were analyzed. (***)p ≤ 0.001 from one-way ANOVA with Tukey’s post hoc test. (C) Macrophages expressing the different constructs were infected with BCG-DsRed at the indicated time points. Cells were lysed and CFU determined. Data represent the Mean ± S.E.M of the CFU recovered compared with control cells (expressing EYFP) from three independent experiments. ns = not significant p > 0.01(*), (*)p ≤ 0.05 from two-tailed Student’s t-test.
