Fig. 6.

Polγ deficiency-mediated autophagy is RICTOR-dependent. a Western blotting shows the overexpression of Rictor or Raptor following transfection of the corresponding vectors in JB6 cells. AKT phosphorylation at Ser473 in the presence of the overexpressed Rictor was also detected by western blotting. Plasmid vector without any protein expression sequence was used as control and is designated by EV. b Detection of LC3 punctation following overexpression of Rictor or Raptor. Quantification of LC3 punctation in Raptor- and Rictor-overexpressing JB6 cells is shown (bottom). c Quantification of LC3 punctation in wild-type MEF and Rictor-knockout MEF cells following Polγ siRNA transfection. d Effects of Polγ deficiency in MEF and Rictor-knockout cells are achieved by expression of Polγ siRNA. The endogenous autophagy markers (LC3 II, beclin 1, ATG7, and p62) were detected by western blotting. The right panels show the corresponding quantification. e Overexpression of Rictor or Raptor in Rictor-knockout MEF cells and the level of LC3 II were determined by western blotting. AKT phosphorylation at Ser473 induced by Rictor overexpression was also detected by western blotting. In all panels, each experiment was repeated at least three times, and statistical analysis was performed using one-way ANOVA analysis followed by Bonferroni’s post-test for multiple-group comparisons. Statistical significance is indicated by asterisks: *p < 0.05 and **p < 0.01