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Fig. 6 Involvement of transcription factors FoxO1 and FoxO3a in NRP-1-mediated EC survival signaling. Phosphorylation of FoxO1 and FoxO3a in NRP-1 survival signaling. After EGF stimulation of EGNP-1-transduced HUVEC or VEGF stimulation of PAEC/NRP-1 for the indicated times, the protein levels in cell extracts of FoxO1 and FoxO3a were assessed by immunoblotting. b. PI3K/Akt depletion resulted in reduction of FoxO1 and FoxO3a phosphorylation. HUVEC were transfected with EGNP-1, then treated with Ly294002 or co-transfected with EGNP-1 and Akt(DN). Immunoblotting was performed to determine the phosphorylation level of FoxO1 and FoxO3a with β-Actin as a loading control. phospho-FoxOs and FoxOs were quantified as a ratio to the amount of β-Actin. *p<0.001, **p<0.005, ***p<0.01, ****p<0.05 in a Student's t test.