1. Effects of ICER deficiency on MB-cAMP-induced activation of PKA-CREB-VEGF signaling and angiogenesis.
Effects of ICER deficiency on MB-cAMP-induced activation of PKA-CREB-VEGF signaling and angiogenesis. Diabetic SIECs were rendered ICER1 deficient (ICER1 siRNA) by using siRNA. Transfection efficiency was confirmed using TaqMan real-time PCR and western blotting. After culturing for 48 hours, cells were exposed to MB-cAMP (100 μM) for various time intervals and were then used to evaluate pCREB-DNA binding activity (Aa, MB-cAMP for 2 hours), CRE transcriptional activity (Ab, MB-cAMP for 6 hours), mRNA expression of CRE target genes (Ba-c, MB-cAMP for 2 hours), HIF-1α-HRE binding affinity (C, MB-cAMP for 2 hours), VEGF promoter activity (Da, MB-cAMP for 24 hours), VEGF mRNA (Db, MC-cAMP for 6 hours) and VEGF protein expression (Dc, MB-cAMP for 24 hours). ‘C-ECs’, control sponge implant endothelial cells; ‘D-ECs’, diabetic sponge endothelial cells. Results are expressed as means±s.e.m. for three independent experiments. *Significantly different from corresponding control values at P≤0.05. **Significantly different from corresponding MB-cAMP-treated control values at P≤0.05. ∫Significantly different from corresponding D-ECs MB-cAMP-treated siRNA control values at P≤0.05.
Milad S. Bitar, and Fahd Al-Mulla Dis. Model. Mech. 2015;8:65-80
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