Fig. 2

E2-induced activation of pERK and its effects on TGFβ1 and TGFβ2 transcription and translation in vitro. a Representative immunoblot of extracellular signal-regulated kinase 1/2 (ERK1/2) activation 1 h post E2 stimulation in vitro. b Densitometry of p-ERK1/2 to total ERK1/2 ratio. c Representative immunoblot and densitometry of p-ERK1/2 to total ERK ratio after 1 h pre-treatment with the mitogen-activated protein kinase (MAPK)/ERK inhibitor, U0126, and subsequent 1 h of E2 stimulation. d, e Steady-state transcript levels of TGFβ1 (d) and TGFβ2 (e) in primary human dermal fibroblasts following a 1 h pre-treatment with U0126 and subsequent 24 h post E2 stimulation. Representative immunoblot of TGFβ1 (f) in primary human dermal fibroblast lysates during a 1 h pre-treatment with U0126 and subsequent 48 h post E2 stimulation. Ethanol, ETOH; estradiol, E2; glyceraldehyde 3-phosphate dehydrogenase, GAPDH; dimethyl sulfoxide, DMSO. Normalized to GAPDH (b, c, f) or B2M (d, e). Bars = mean ± SEM. Data shown is from ≥ 6 independent experiments using dermal fibroblasts from ≥ 6 different donors. Statistical analysis: Two-tailed, Wilcoxon matched-pairs signed rank test (b) and one-way ANOVA with Sidak’s multiple comparisons post hoc test (c–e). *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.005, ****p ≤ 0.0001