Cleaved-Caspase 4 (Gln81) Antibody - #AF5373

Figure 4. Melatonin attenuates DON-induced apoptosis by suppressing ER stress in human granulosa cells. (A) Cells were treated with different concentrations of DON for 24 h. The protein levels of BiP, CHOP, and phosphorylated-eIF2α were assessed by western blot. (B) For the inhibition of ROS production, Mito-TEMPO was added 1 h before DON treatment. Cells were cultured for another 24 h and the levels of BiP, CHOP, and phosphorylated-eIF2α were measured by western blot. (C) Granulosa cells were cultured in medium containing DON with or without melatonin for 24 h. The levels of BiP, CHOP, and phosphorylated-eIF2α were analyzed by western blot. (D) The levels of Cleaved Caspase 4 and Phosphorylated JNK were analyzed by western blot. (E) ER stress inhibitor 4-PBA was added 1 h before DON and melatonin incubation. The cells were cultured for 24 h and cell viability was determined using MTT assay kit. (F) The protein levels of Bax, Bcl-2, and Cleaved Caspase-3 from granulosa cells treated as depicted in Figure 4D were analyzed by western blot. (G) Quantification of Bcl-2 and Bax expression using densitometric analysis. β-actin served as the control for loading. Data are represented as means ± SEM of three independent experiments. ∗P < 0.05, ∗∗P < 0.01.