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. Author manuscript; available in PMC: 2018 Apr 1.
Published in final edited form as: Gynecol Oncol. 2017 Jan 29;145(1):167–175. doi: 10.1016/j.ygyno.2017.01.022

Figure 3. FVIIa stimulates VEGF-A release from human EOC cell lines in a PAR-2 dependent manner.

Figure 3

(A) Whole cell lysates prepared from control HEK-293T (epithelial) and EA.hy926 (endothelial) cell lines; and EOC cell lines SKOV-3, OVCAR-3 and CaOV-3 were separated by 10% SDA-PAGE and immunoblotted with anti-PAR-2 (upper panel) and β-actin loading controls (lower panel). (B) Relative VEGF-A concentration in serum free media 48 hours after exposure of SKOV-3 cell line to FVIIa (0–50 nM) was quantified. VEGF-A was detected by ELISA and expressed as fold increase over baseline. (C) Comparison of fold increase in VEGF-A concentration in response to FVIIa (50nM) for EOC cell lines SKOV-3, OVCAR-3 and CaOV-3 at 48 hr (first bar- basal levels). (D) Dose dependent inhibition of FVIIa-induced VEGF-A release at 48 hr in SKOV-3 cells preincubated with the small molecule PAR-2 antagonist ENMD-1068 (0.01–1 mM). (E) Inhibition of FVIIa induced VEGF-A release in (i) SKOV-3, (ii) OVCAR-3 and (iii) CaOV-3 cells at 48 hrs by preincubation with 1 mM PAR-2 antagonist (ENMD-1608). All results are expressed as the mean fold increase in VEGF-A concentration (n=3) with error bars representing ± S.D. (* p<0.05, ** p<0.001).