GPR30 activation attenuates TNF-mediated endothelial expression of ICAM-1 and VCAM-1.

<p>(A and B) Confluent HUVEC monolayers were pre-treated with the selective GPR30 agonist G-1 (0.1–3 µM) for 45 minutes prior to 4 hour stimulation with TNF (5 ng/ml). Cells were lysed and the lysates were immunoblotted with antibodies against ICAM-1 (A), VCAM-1 (B) and α-tubulin. DMSO (1∶5000) was used as a solvent control for G-1. (C and D) Confluent HUVECs were pre-treated for 30 minutes with/without the specific GPR30 inhibitor G-15 (2 µM) followed by treatment with G-1 (1 µM) for 45 minutes prior to 4 hour stimulation with TNF (5 ng/ml). Cells were lysed and the lysates were immunoblotted with antibodies against ICAM-1 (C), VCAM-1 (D) and α-tubulin. Data shown are mean ± SEM of 6–8 independent experiments. *, ** and *** indicate p<0.05, p<0.01 and p<0.001 respectively, compared to the untreated control.