Fig. 6

VCP is required for autophagy initiation signaling in HCN cells following insulin withdrawal. a-b Negative regulators of autophagy, mTOR and calpain 2 were suppressed in I(-), but reactivated by DBeQ treatment (a) or VCP knockdown (b). On the contrary, positive inducers of autophagy such as GSK-3β were activated following insulin withdrawal, but deactivated by DBeQ treatment (a) or VCP knockdown (b). c The high number of DFCP1 puncta observed in I(-) were greatly reduced by DBeQ treatment. The number of DFCP1 puncta was quantified with cut-off diameter of 0.5 μm. Scale bar, 5 μm. Quantitative DFCP1 puncta were determined as the mean ± SD by One-way ANOVA followed by Tukey’s multiple comparison test (n = 10). ***p < 0.001