Critical role of S1PR2 in endothelial inflammation in vitro. (A) S1PR1, S1PR2, and S1PR3 mRNA levels in HUVEC. HUVEC were incubated with vehicle (–), TNF-α, or basic fibroblast growth factor (bFGF) in 0.5% fetal bovine serum endothelial basal media-2 for 12 hours. Shown are the results of quantitative real-time PCR analyses (fold induction TNF-α–treated vs non-treated cells). Data are mean ± SEM of three independent experiments. *P < .05 treated vs non-treated cells. (B-F) mRNA levels of E-selectin, VCAM-1, ICAM-1, TF, and MCP-1 in HUVEC. HUVEC were incubated with vehicle (V) or TNF-α (TNF) in the absence or presence of the S1PR2-specific antagonist JTE013 (JTE), the S1PR1-specific antagonist W146, or the S1PR1 and S1PR3 antagonist VPC 23019 (VPC). Shown are the results of quantitative real-time PCR analyses (fold induction TNF-α–treated vs non-treated cells). (B-G) All S1PR antagonists were used at 30 μM concentration. (G) Blockade of S1PR2 signaling in HUVEC inhibits U937 monocyte adhesion. HUVEC were treated with vehicle (V) or TNF-α (TNF) in the absence or presence of the S1PR2-specific antagonist JTE013 (JTE), the S1PR1-specific antagonist W146, or the S1PR1 and S1PR3 antagonist VPC 23019 (VPC). Results are mean ± SEM of quadruplets of 1 representative experiment of 3 experiments with similar results. *P < .05 TNF-α–treated vs non-treated cells and, where indicated, between TNF-α–treated and TNF-α+ antagonist–treated cells.