Figure 4. 25-Hydroxycholesterol (25-HC) protected PC12 cells from I/R-induced cytotoxicity and inhibits autophagy following oxygen-glucose deprivation/reperfusion (OGD/R) in PC12 Cells. (A) PC12 cell viability is assessed by measuring CCK8. Results were shown as fold of control. (B–F) PC12 cells were pretreated with 25-HC and then cultured under OGD/R conditions or normoxia. Apoptosis analysis was performed, and representative flow cytometry images are shown. Quantifications of the percentage of apoptotic cells are shown (N = 3 per group, ***P<0.001, **P<0.01 versus sham group, ##P<0.01, #P<0.05 versus OGD/R group). (G) The expression of cleaved caspase3 was tested by immunocytofluorescent staining (N = 3 per group, ***P<0.001 versus sham group, ###P<0.001 versus OGD/R group). (H) The expressions of cleaved caspase-3, caspase-3, Bcl-2, and Bax were measured by Western blot (N = 3 per group, **P<0.01, *P<0.05 versus sham group, #P<0.05, versus OGD/R group).