CRAMP prevented PDGF-BB-enhanced ROS by targeting NOX1. (A) VSMC were pretreated with CRAMP (100 ng/mL) for 2 h and then stimulated with PDGF-BB (20 ng/mL) at different times (0.5, 1, 2, 4, 6 h). The level of ROS was detected using fluorescence microplate reader. Data of 3 independent experiments is presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with control, n = 5. (B) VSMC were pretreated with CRAMP (100 ng/mL) for 2 h and then stimulated with PDGF-BB (20 ng/mL) for 4 h. The level of ROS was detected by immunofluorescence detection. (C) VSMC were pretreated with CRAMP (100 ng/mL) for 2 h and then stimulated with PDGF-BB (20 ng/mL) for 24 h following by immunoblotting with anti-NOX1, anti-NOX2 and anti-NOX4 antibodies. (D) VSMC were pretreated with CRAMP (100 ng/mL) for 2 h and then stimulated with PDGF-BB (20 ng/mL) for 24 h followed by immunoblotting with anti-α-SMA and anti-SM22α antibody. Data of 3 independent experiments is presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with control, n = 3.