Fig. 4: High expression of endogenous C15ORF48 enhances autophagy.

a A549 and MDA-MB-231 cells were lysed and subjected to western blotting with the indicated antibodies. Data are representative of three independent experiments with three biological replicates. b, c As in Fig. 2b, c, except that A549 and MDA-MB-231 cells were used. Statistical significance was calculated using two-way ANOVA followed by Tukey’s multiple comparisons test (n = 70 cells from two independent experiments). d MDA-MB-231 cells were transfected with the indicated siRNAs for 48 h. After incubation, cells were used for ATP assays. ATP amount in a cell is shown as means ± SDs. Statistical significance was calculated using two-way ANOVA followed by Tukey’s multiple comparisons test (n = 3, biological replicates). e MDA-MB-231 cells were transfected with the indicated siRNAs for 48 h. 44 h after transfection, media were replaced with fresh media. After incubation, cells were lysed and subjected to western blotting, as in Fig. 1d. f, g MDA-MB-231 cells were transfected with the indicated siRNAs for 48 h. 44 h after transfection, media were replaced with fresh media. Half the samples were treated with bafilomycin A1 (Baf A1, 200 μM) for the final 1 h. Cells were fixed and analyzed for LC3 puncta by immunocytochemistry, as in Fig. 2b, c. Statistical significance was calculated using two-way ANOVA followed by Tukey’s multiple comparisons test (n = 70 cells from two independent experiments).