GRP78 overexpression induced autophagy activation in PC12 cells. (A), (B): The control, LV-vector, LV-GRP78-PC12 cells were harvested and subjected to Western blotting analysis. (A) LC3II/LC3I was upregulated in LV-GRP78-PC12 cells. (B) Beclin1 was upregulated in LV-GRP78-PC12 cells. (C) Electron microscopic images show increased number of double-membrane vacuolar structures in the LV-GRP78-PC12 cells. The cells were fixed in 0.25% glutaradehyde and examined with a transmission electron microscope. N: nucleus. Arrows indicated typical autophagosomes. Scale bar = 100 nm. (D), (E) LV-GRP78-PC12 cells were transfected with GRP78 siRNA1 (40 nM) or negative control sequence (NC). Forty-eight hours later, the cells were harvested and subjected to Western blotting analysis. (D) GRP78 siRNA1 reduced GRP78 expression. (E) GRP78 siRNA1 reduced LC3II/LC3I. Bar represents mean ± SD, n = 3. **P < 0.01, ***P < 0.001 compared with LV- vector; # P < 0.05, ### P < 0.001 compared with NC.