Figure 3.

GR inhibitors restore cortisol-dependent inhibition of collagen type I synthesis and AP collagen peptides inhibit GR phosphorylation. HDFs were treated with cortisol (100 nM) in the presence of an MR inhibitor (spironolactone, 1 μM) or GR inhibitor (RU486, 1 μM) and cultured for 24 h. (A) Total RNA was extracted from cells and relative col1a1 mRNA levels were measured by qRT-PCR. (B) Culture media were harvested, and the level of procollagen type I was detected using a specific ELISA kit. (C,D) Cell lysates from cells cultured for 8 h were subjected to immunoblot analysis to assess the phosphorylation of GR. All data represent the mean ± SD of three independent experiments. Significant differences: ## p < 0.01, ### p < 0.005 compared with the control; * p < 0.05, ** p < 0.01, *** p < 0.005 compared with cortisol-treated control.