PlGF-induced LTE4 release, FLAP mRNA expression, and ROS formation. (A) PBM or THP-1 cells were treated with PlGF (250 ng/mL) for 24hours and (B) THP-1 cells were pretreated for 30 minutes with Ab-VEGFR1 (2 μg/mL), LY294002 (10 μM), DPI (10 μM), MK866 (10 μM), and U73122 (10 μM), followed by treatment with PlGF for 24 hours. The supernatants were collected and assayed for LTE4 release by ELISA. (C) RPA analysis of total RNA isolated from THP-1 cells pretreated for 30 minutes with Ab-VEGFR1, LY294002, DPI, R59949 (10 μM), PDTC (10 μM), SB203580 (10 μM), and PD98059 (10 μM) before PlGF treatment for 24 hours. (D) THP-1 cells were loaded with DCFH-DA dye for 30 minutes, incubated with indicated inhibitors for an additional 30 minutes and treated with PlGF for 4 hours. The cells were lysed, and the fluorescence was measured. Data are expressed as means plus or minus SEM of 3 independent experiments (***P < .001; **P < .01; ns, P > .05). Where indicated, the vertical lines show repositioned gel lanes.
