Specific siRNA knockdown of the Hedgehog signaling pathway and Gas1-mediated autophagy in LX-2 and T-6 cells. A: LX-2 and T-6 cells were transiently transfected with Gas1 siRNA or control siRNA (siR Con) for 72 h, and then the expression levels of key components of the Hedgehog signaling pathway (Patch1 and Gli1), LC3-II and p62 were measured by western blotting. B and C: The downregulation of Patch1 and Gli1 was measured by immunoblotting protein extracts derived from Gas1 siRNA-treated and control siRNA-treated LX-2 and T-6 cells. D and E: The autophagy markers, LC3-II and p62, were assessed after Gas1 siRNA transfected by immunoblotting. Gas1 siRNA promotes autophagic activity in LX-2 and T-6 cells. GAPDH was used as a loading control. The data are presented as the means ± SD for at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, unpaired Student’s t-test.