Figure 2. CAP induces the ER stress response.

(A) The levels of eIF2α phosphorylation at Ser⁵¹ and ATF4 expression were evaluated in MEFs incubated with or without 100 μg/ml CAP for 48 h. The abundance of both phosphorylated and total eIF2α was assessed by Western blotting. Immunodetection of β-actin was used as the loading control. All experiments were done in triplicate. Quantification is shown in the right panel. *P<0.05 **P<0.01, ***P<0.001 vs control. (B) Chop-10 mRNA abundance was evaluated by RT-qPCR in MEFs treated with CAP. Chop-10 mRNA abundance was also evaluated in MEFs treated with the antibiotics, actinonin (50 μM), doxycycline (100 μg/ml) and CAP (100 μg/ml), for 48 h. Chop-10 mRNA was normalized to 18S rRNA, which was the loading control. All experiments were done in triplicate; *P<0.05.