Fig. 3

Genetic inhibition of VCP changed ACD to apoptosis in insulin-deprived HCN cells. a An experimental scheme for the knockdown of VCP. CDA, cell death assay; WB, Western blotting. b Knockdown of VCP was confirmed by Western blotting after 24 h treatment. NT, non-targeting. c VCP knockdown increased cell death and Z-VAD-FMK efficiently blocked an increase of cell death in I(-) HCN cells. Quantitative data are determined as the mean ± SD by One-way ANOVA followed by Tukey’s multiple comparison test (n = 3). ***p < 0.001. n.s., not significant. d Cleaved caspase 3 was detected in I(-) HCN cells following VCP knockdown at 48 h. e Cell death rate was not affected by DBeQ in I(-) HCN cells depleted of VCP. Quantitative data are determined as the mean ± SD by One-way ANOVA followed by Tukey’s multiple comparison test (n = 3). ***p < 0.001. n.s., not significant. f Atg7 knockdown decreased cell death in I(-) HCN cells. However, cell death rate was not altered by DBeQ or Z-VAD-FMK in I(-) HCN cells with stable knockdown of Atg7. Quantitative data are determined as the mean ± SD by One-way ANOVA followed by Tukey’s multiple comparison test (n = 3). ***p < 0.001. n.s., not significant