Fig. 3

GRP78 regulates autophagy via AMPK/mTOR in A2E and blue light-treated RPEs. a: Wild RPEs and RPEs received GRP78 shRNA (GRP78i) or negative control shRNA (NCi) were treated with 25 μM A2E and blue light exposure. Cells were collected 12 h later. GRP78, AMPK, mTOR, p-AMPK, p-mTOR and LC3 protein levels were assessed by western blot analysis. Bars represent the mean ± SD from three independent experiments (n = 3, *p < 0.05). b: LC3 immunofluorescence was detected at 12 h after treatment. In wild RPEs or in negative vector group, LC3 revealed an increased dots distribution. GRP78 shRNA attenuated the formation of LC3 puncta in A2E and blue light-treated RPEs (scale bar, 10 μm). The number of autophagosomes was obtained by counting LC3 fluorescent puncta in RPEs from three independent experiments (n = 3, *p < 0.05).