Table 4.

Inhibition of Akt-induced kinase insert domain protein receptor and neuropilin-1 mRNA expression by signaling inhibitors

Transcript name	Infection with	After infection	+Wortmannin	+U0126	+SB203580	+Wortmannin/SB203580
Kinase insert domain protein receptor	Ad-βgal	+1.7	+1.2	-1.1	+1.7	+1.3
	Ad-Akt	+3.2*	+1.6	+1.4	+2.3*	+1.5
Neuropilin-1	Ad-βgal	+1.5	+1.1	+1.6	+1.7	+1.3
	Ad-Akt	+2.6*	+1.3	+1.6	+2.4*	+1.1

EPCs were treated with 200 nM Wortmannin (a phosphatidylinositol-3 kinase inhibitor), 10 μM U0126 (a mitogen-activated protein kinase [MAPK]/extracellular signal-regulated kinase [MEK]/Erk-1/2 inhibitor), or 25 μM SB203580 (a P-38 MAPK inhibitor). One hour later, EPCs were infected with β-galactosidase (Ad-βgal) or myr-Akt (Ad-Akt), and the mixture was incubated for 24 h. Total RNA was then harvested for analysis of VEGFR-2- and NRP-1 mRNA expression. In order to standardize the quantitation of kinase insert domain protein receptor and neuropilin-1, glyceraldehydes-3-phosphate dehydrogenase (GAPDH) from each sample was quantified by QRT-PCR and kinase insert domain protein receptor and neuropilin-1 were normalized to GAPDH. Values represent fold change (FC) comparing kinase insert domain protein receptor and neuropilin-1 after infection with β-galactosidase (Ad-βgal) or myr-Akt (Ad-Akt) and coincubation with the indicated inhibitors compared to control. All analysis values represent SEM performed in triplicates

^*P < 0.05