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. Author manuscript; available in PMC: 2016 Jan 6.
Published in final edited form as: Circ Res. 2015 Jan 21;116(7):1120–1132. doi: 10.1161/CIRCRESAHA.116.304408

Figure 5. PDE1C regulates PDGFRβ protein levels via a cAMP-PKA dependent mechanism.

Figure 5

(A) PDE1 inhibitor IC86340 dose-dependently decreased PDGFRβ protein levels. Rat aortic SMCs were treated with indicated doses of IC86340 for 24 h. (B) Knockdown of PDE1C but not PDE1A decreased PDGFRβ protein. Rat aortic SMCs were transfected with adenoviral vectors expressing a high dose of adenovirus encoding LacZ-shRNA, PDE1A-shRNA, or PDE1C-shRNA for 3 days. (C) Forskolin enhanced the effect of IC86340 on PDGFRβ protein reduction. Rat aortic SMCs were treated with a low dose of IC86340 (5 μmol/L) with or without 10 μmol/L forskolin for 24 h in DMEM containing 0.1% FBS. (D) Forskolin augmented PDE1C knockdown-induced PDGFRβ protein reduction. Rat aortic SMCs were transfected with a low dose of adenovirus encoding LacZ-shRNA, PDE1A-shRNA, or PDE1C-shRNA for 3 days, followed with or without 10 μmol/L forskolin treatment for 24 h in DMEM containing 0.1% FBS. (E) PKA inhibitor H89 blocked IC86340-induced PDGFRβ protein reduction. Rat aortic SMCs were treated with a high dose of IC86340 (15 μmol/L) with or without of 5 μmol/L H89 for 24 h in DMEM containing 0.1% FBS. (F) Specific PKA inhibitor PKI (14-22) blocked IC86340-induced PDGFRβ protein reduction. Rat aortic SMCs were treated with a high dose of IC86340 (15 μmol/L) with or without 5 μmol/L PKI (14-22) for 24 h in DMEM containing 0.1% FBS. PDGFRβ protein levels and b-actin equal loading were analyzed by immunoblotting with the anti-PDGFRβ and β-actin antibody, respectively. The blots were analyzed by densitometry. Fold changes normalized to the left lane. Values are mean ± SD (n= 3). *p<0.05.

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