Fig. 1.
Platelet-derived growth factor (PDGF)-induced repression of CPI-17 gene. A: total protein extracts (20 μg) and total RNA preparation (0.2 μg) of rat aorta tissues and the cell culture [rat aorta smooth muscle cell (AoSMC)] were subjected to immunoblotting (left) and quantitative (q)RT-PCR (right), respectively. CPI-17 protein extent was normalized against GAPDH, and the ratio in aorta tissues was set as 1 (n = 6). Relative extent of CPI-17 mRNA was obtained by ΔΔCt method using GAPDH as an internal control (n = 9). B: subconfluent AoSMC was incubated for overnight with MCDB medium supplemented with 1% FBS. Quiescent cells were restimulated for 24 h with 10% FBS or 50 ng/ml PDGF-BB, subjected to qRT-PCR analysis, using β-tubulin as an internal control (n = 6). C: quiescent AoSMC was stimulated with 50 ng/ml PDGF or 1 μM ANG II for 24 h in the presence of kinase inhibitors (10 μM U1026, 10 μM SP600125, and 10 μM SB203580), which are added 30 min before the stimulation (n = 3–6). **P < 0.05 and #P < 0.05, compared with unstimulated and no-inhibitor controls, respectively.