Figure 3.

Effects of autophagic flux dysregulation promoted by ATP6V1B2 and ATP6V1C1 mutants

Confocal laser scanning microscopy analyses show a significant storage of cholesterol (A and B) and ceramide (C and D) on patients’ fibroblasts compared with control cells. Cells were stained with filipin (cholesterol) (A) and BTR-ceramide (C) probes, respectively. Scale bars are 10 μm (A), 20 μm (C, left), and 2 μm (C, right). Arrows indicate the selected cells reported in the enlarged images. MFI ± SEM of filipin (B) and ceramide (D) signal per cell, was quantified (3 independent experiments, ≥20 cells per condition, in each repeat), and plotted in the corresponding graphs, defining Final MFI as MFI (region of interest) – MFI (background). WT1 and WT2 were pooled together vs. patient’s samples. Bars indicate mean ± SEM, p values were calculated by One way ANOVA with Tukey’s correction for multiple testing.