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. 2009 Nov 4;42(1):21–29. doi: 10.3858/emm.2010.42.1.002

Figure 3.

Figure 3

Effect of SHP on angiotensin II-stimulated AP-1 activity. (A) Luciferase gene reporter assay of the effect of SHP on Smad3-independent PAI-1 promoter activity. VSMCs were transfected with a luciferase reporter gene under the control of an SBE deleted PAI-1 promoter (-240 PAI-1 promoter Luc, 300 ng), together with the indicated amounts of SHP expression vector for 5 h. Cells were incubated in medium containing 0.5% FBS for 24 h, and then treated with angiotensin II (Ang II, 10 nM) for 1 h before harvest. Data represents the means ± SEM. *P < 0.001 as compared to control. **P < 0.01 as compared to Ang II alone (WT PAI-1 promoter), #P < 0.05 as compared to Ang II alone (SBE-deleted PAI-1 promoter). (B) Electrophoretic mobility shift assay of the effect of SHP on AP-1 DNA binding activity. VSMCs were infected at the indicated moi with Ad-SHP or GFP for 4 h. Cells were incubated in medium containing 0.5% FBS for 24 h, and then treated with angiotensin II (Ang II, 10 nmol/l) for 1 h before harvest. (C) VSMCs were cotransfected with the -840 PAI-1 promoter Luc (300 ng) and the AP-1 decoy ODN (10 nM) or mismatched AP-1 decoy ODN (MD, 10 nM) together with the indicated amounts of an SHP expression vector (pcDNA3) or c-jun/c-fos expression vector for 5 h. Cells were incubated in medium containing 0.5% FBS for 24 h, and were then treated with angiotensin II (Ang II, 10 nmol/l) for 1 h before harvesting. Data represents the means ± SEM. *P < 0.05, ##P < 0.001 as compared to the control. **P < 0.05, #P < 0.01 were as compared to Ang II.