Analysis of S1PR subtype contribution to S1PR agonist-induced ECM synthesis.
A, relative S1PR mRNA expression in NHLF was analyzed by qPCR. The data show the means ± S.E. of three independent experiments. For normalization, 18 S, HPRT1, and PPIA were used. B, NHLF were preincubated with vehicle, S1P1R, S1P2R, or S1P3R antagonist or a combination of S1P2R and S1P3R antagonists for 1 h before stimulation with S1PR agonists (5 μm) for 24 h. ECM synthesis was then measured with the [3H]proline incorporation assay. The data represent the means ± S.E. of three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (t test, vehicle versus antagonist treatment).