Fig. 8.

FCPR03 reduces intracellular ROS and represses ER stress in OGD-treated HT-22 cells. (A, B) HT-22 cells pretreated with FCPR03 (20 μM) or N-Acetyl-l-cysteine (NAC, 2.5 mM) for 1 h were subjected to 6 h of OGD and were incubated with CellROX Deep Red Reagent (5 μM) for 30 min. Finally, intracellular ROS was detected by confocal microscopy. Intracellular ROS was quantified by Image J (18 fields from three independent experiments, n = 3). (C) HT-22 cells pretreated with FCPR03 or NAC for 1 h were subjected to 6 h of OGD. Lipid peroxidation in HT-22 cells was measured by the level of malondialdehyde (MDA). (D, E, F, G) HT-22 cells pretreated with NAC for 1 h were subjected to 6 h of OGD. The protein levels of GRP78, p-eIF2α, and eIF2α were determined by Western blotting. The relative levels of GRP78/β-tubulin and p-eIF2α/eIF2α were determined by semiquantitative analysis of the Western blots (n = 3). (H) HT-22 cells pretreated with FCPR03 or NAC for 1 h were subjected to OGD for 6 h and re-oxygenation for 24 h, and then cell viability was detected by a CCK-8 assay (12 duplications from three independent experiments, n = 3). Results are expressed as the mean ± SD, *P < 0.05, **P < 0.01 versus the indicated group. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)