Figure 5.

Gas1 elevates autophagic activity in the presence of Hedgehog pathway antagonists. A: Autophagy was induced in LX-2 and T-6 cells by culturing in EBSS for 4 h, cells were then were treated with cyclopamine (5 µM) for 72 h. Endogenous Patch1, Gli1, LC3B and p62 were detected by immunoblotting of total cell lysates, quantified by densitometric analysis and normalized to GAPDH. B and C: Patch1 and Gli1 expression levels were determined by western blotting. Patch1 and Gli1 were downregulated in LX-2 and T-6 cells treated with cyclopamine(Cyc, 5 µM), compared with the negative control group. D and E: Endogenous LC3-II and p62 were detected in cell lysates from cells treated with cyclopamine for 72 h. The results shows that Gas1 promotes the autophagic activity in LX-2 and T-6 cells. Quantification by densitometric analysis relative to GAPDH expression is shown on the graph. The results are representative of three independent experiments. Bars represent means ± SD. *P<0.05, **P<0.01, ***P<0.001 versus the negative control group, Unpaired student’s t-test.