Figure 4. Effect of ATF4 silencing on CAP-induced ISR gene expression.

(A) MEFs stably transfected with either non-targeting shRNA (shGFP) or with two different shRNAs against ATF4 (shATF4) for 24 h was replaced with control medium or with medium containing CAP (100 μg/ml) for 24 h. CAP induced CoxI decline were assessed by Western blotting. Immunodetection of GAPDH was used as the loading control. All experiments were done in triplicate. Quantification is shown in the right panel.*P<0.05, ***P<0.005 vs control. (B) MEFs which were stably transfected were incubated with or without CAP (100 μg/ml) for 12 h. The mRNA expression of ISR and mtUPR-related genes was evaluated by RT-qPCR. Results are expressed as fold change to control and are expressed the mean ± SD of three independent experiments. ***P<0.005 vs. control.