Fig. 7

CRP2 regulates Ang II-induced MMP2 and Col III via Erk1/2 signaling. A Apoe^−/− VSMCs were pretreated with U0126 for 30 min before Ang II stimulation. Proteins or conditioned medium were prepared 24 h later. Western blot analysis was performed to detect Erk1/2 phosphorylation, total Erk1/2, CRP2, MMP2, and Col III expression. α-Tubulin was used to verify loading. Zymography was performed to measure MMP2 activity. A representative of 3–4 independent experiments is shown. B Quantitation of MMP2 activity (n = 4), MMP2 (n = 4) and Col III (n = 3) levels. The expression level of vehicle-treated, without Ang II stimulation was set as 100%. §P < 0.05 compared with the vehicle baseline without Ang II; *P < 0.05 compared with the corresponding vehicle group. C Left panel, Csrp2^−/−Apoe^−/− VSMCs were transfected with pMyc-His or pMyc-His-CRP2 expression plasmids and treated with Ang II. Conditioned medium and total proteins were then collected 24 h later for zymography to determine MMP2 activity and protein expression (Myc, CRP2, and α-tubulin), respectively. A representative of 3 independent experiments is shown. Right panel, Quantitation of MMP2 activity and expression level. pMyc-His-vector-transfected MMP2 activity or expression level without Ang II stimulation was set as 1. n = 3 each group. §P < 0.05 compared with the vector baseline without Ang II; *P < 0.05 compared with the corresponding vector group. D Csrp2^−/−Apoe^−/− VSMCs were transfected with pEGFP or pEGFP-CRP2 plasmids and treated with Ang II. After 24 h, conditioned medium was collected for zymography to determine MMP2 activity and total proteins prepared for Western blot analysis to detect expression levels of MMP2, Col III, CRP2-EGFP, and α-tubulin as a loading control