MRGPR-X1 induce EGR-1 via ERK-1/2 in F11-MRGPR-X1 cells.

<p>RTQ-PCR experiments were performed with cDNAs derived from serum-starved F11-MRGPR-X1 cells stimulated or not with BAM8-22 (1 µM) for 1 h (A, C and D) using specific primers for 15 distinct genes as indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0058756#pone-0058756-t001" target="_blank">table 1</a>. The name of the analyzed gene is listed beneath the corresponding bar. Genes with a p value <0.05 are show in black bars. In (C and D) cells were treated or not with PD-184352 (30 min, 10 µM). Relative BAM8-22-induced gene expression was normalized to β-actin and calculated using the ΔΔCp method. In (B) serum-starved F11-MRGPR-X1 cells were stimulated or not with BAM8-22 (1 µM) for the indicated period of time and expression of EGR-1 was determined by western-blotting. Afterwards blots were stripped and re-probed with an antibody against ERK-2 (t-ERK-2). One representative blot is shown. Ligand-induced EGR-1 expression was quantified by densitometry and is given normalized to not stimulated cells. Data from 4 independent experiments were compiled and expressed as the mean ± S.E.M. Asterisk indicate a significant difference to not stimulated cells. Dagger signs indicate a significant difference between BAM8-22-stimulated inhibitor-treated and untreated cells.