Fig. 4

VCP inhibition reduced autophagic flux in insulin-deprived HCN cells. a-b DBeQ treatment (a) or VCP knockdown (b) induced a moderate increase in LC3-II in I(+) HCN cells. Blocking of autophagy using BafA1 gave rise to a significant increase of LC3-II with the same levels between BafA1 alone and BafA1/DBeQ or BafA1/VCP knockdown cells in I(+). Quantitative LC3-II levels were determined as the mean ± SD by One-way ANOVA followed by Tukey’s multiple comparison test (n = 3). **p < 0.01, ***p < 0.001. n.s., not significant. c-d DBeQ treatment (c) or VCP knockdown (d) led to a decrease in LC3-II in I(-) HCN cells. Blocking of autophagy using BafA1 gave rise to a significant increase of LC3-II, but the amount of accumulated LC3-II in BafA1/DBeQ or BafA1/VCP knockdown cells was significantly less than BafA1 alone in I(-). Quantitative LC3-II levels were determined as the mean ± SD by One-way ANOVA followed by Tukey’s multiple comparison test (n = 3). **p < 0.01, ***p < 0.001. n.s., not significant. e mRNA expression level of LC3 did not show different changes by VCP inactivation in I (-). Quantitative LC3 mRNA expression levels are represented as the mean ± SD (n = 3). Quantitative data was determined by Student’s t-test. *p < 0.05. n.s., not significant