Figure 8. GSK3 inhibitor, CHIR-99021, rescues the expression of autophagy-related proteins and nuclear receptor GR, RORα expression in light exposure cells. (A–B) Protein content of LC3B, Beclin, SIRT1 and phospho-AMPK. GSK3 inhibitor CHIR-99021 completely rectified light-induced up-regulation of autophagy-related proteins, including LC3B, Beclin, SIRT1 and phospho-AMPK, in HT-22 cells. Values are means ± SEM, **p < 0.01, compared with Dark group, ##p < 0.01, compared with Light group (n = 4); (C–E) Immunofluorescence of GR, showing that CHIR-99021 was able to restore light-induced GR activation and GR nuclear translocation. The nuclei were stained with Hoechst (blue) and GR was stained with GR antibody (red). Scale bars, 75 μm; (F–G) Protein content of GR, phospho-GR, RORα and REV-ERBα. CHIR-99021 significantly alleviated light-induced increase of phospho-GR and RORα protein expression, yet the increase of total GR and the decrease of REV-ERBα in Light group were not restored. Values are means ± SEM, *p < 0.05, **p < 0.01, compared with Dark group, #p < 0.05, ##p < 0.01, compared with Light group (n = 3 or 4).