Fig. 3

BCRP3 promotes PI3P production but does not affect TORC1, AMPK, and ULK1 activities. A, C Immunofluorescence staining of ATG16L1 in control or BCRP3-deficient HeLa cells (A) or HeLa cells stably expressing vector or BCRP3 (C) and starved in EBSS for 2 h. Representative confocal images are shown on the left and quantitative data are on the right. Arrowheads indicate the ATG16L1 puncta. Bar, 10 μm. Data are means ± SD from three independent experiments and 30 cells per group per experiment were counted. P values are determined by one-way ANOVA with Tukey’s post hoc test (A) or unpaired t-test (C), *P < 0.05, **P < 0.01. B, D Confocal microscopy analysis of HeLa cells derivatives as in (A) or (C), respectively, transiently transfected with GFP-DFCP1, and starved in EBSS for 2 h. Representative images are shown on the left and quantitative data are on the right. Bar, 10 μm. Data are means ± SD from three independent experiments and 20 (B) or 10 (D) cells per group per experiment were counted. P values are determined by one-way ANOVA with Tukey’s post hoc test (B) or unpaired t-test (D), ***P < 0.001. E, F Western blot analysis of indicated proteins in control or BCRP3-deficient HeLa cells (E) or HeLa cells stably expressing vector or BCRP3 (F) and starved in EBSS for 1 h. The blots are representative of n = 3 independent experiments. Quantitative data are shown on the right, ns, not significant by one-way ANOVA with Tukey’s post hoc test (E) or unpaired t-test (F)