Downregulation of c-Jun/Ap-1 by c-Jun-siRNA suppresses Chop expression. (A) mRNA levels of c-Jun were assessed by RT-qPCR in MOVAS cells transfected with siRNAs for 24 h; GAPDH was used as an internal control (n≥3). (B) Western blotting demonstrated the Ap-1 levels in MOVAS cells transfected with siRNAs for 48 h; GAPDH was used as an internal control (n=3). MOVAS cells were transfected with siRNAs for 24 h and the transfected cells were treated with Ang II (20 nM) for 24 h. (C) The mRNA expression level of Ddit3 were detected by RT-qPCR; GAPDH was used as an internal control (n≥3). (D) Protein levels were detected by western blotting with anti-c-Jun, anti-Chop, and anti-β-actin (internal control) antibodies (n=3). All results are presented as the mean ± standard error of the mean. #P<0.05; *P<0.05, **P<0.01, ***P<0.001 vs. NC-si. Ap-1, activator protein 1; Chop, C/EBP homologous protein; RT-qPCR, reverse transcription quantitative-polymerase chain reaction; siRNA, small interfering RNA; Ang II, angiotensin II; NC-si, negative control siRNA; Ddit3, DNA damage-inducible transcript 3 protein.