Figure 4.

Contribution of MEK/ERK to cAMP regulation of CCND1 and IGFBP1 expression in cultured endometrial stromal fibroblasts. A–C, Endometrial stromal fibroblasts from patients without (no endo, gray bars, n = 9) or with (endo, white bars, n = 7) endometriosis were treated with DMSO (vehicle, no inh) or 10 μm U0126 for 30 min in 2% serum-containing medium, followed by addition of vehicle (Ctrl) or 0.5 mm 8-bromo-cAMP for 4 d. Treatments and medium were refreshed every 2 d. CCND1 (A) and IGFBP1 (B) mRNA expression was analyzed by quantitative RT-PCR, and values were normalized to the RPL19 gene. C, IGFBP1 protein secretion was analyzed by ELISA, and values were normalized to total RNA. All bar graphs (least square means ± sem) represent fold changes relative to control (Ctrl, no endo and no treatment). Means with same letters indicate significant differences within treatments at P < 0.05 using two-way ANOVA by ranks, followed by Bonferroni post hoc test. Means with asterisks indicate significant differences in endo vs. corresponding no endo treatment at P < 0.05 using Mann-Whitney U test.