Ang II-induced collagen I gene
(-)-MK 801 in dermal fibroblasts. (A) Effects of actinomycin D and cycloheximide on Ang II-induced proα1(I) mRNA expression. Cells were preincubated for 30 min with actinomycin D (10 μg/ml) or cycloheximide (10 μg/ml), followed by treatment with or without Ang II (1 μM) for 24 h. A representative Northern blot analysis of four experiments is shown. (B) Nuclear run-on analysis. Cells were incubated with or without Ang II (1 μM) for 24 h. Nuclei were isolated, and 32P-labeled nuclear RNA transcripts were hybridized to proα1(I) or GAPDH. A representative autoradiogram of four independent experiments is
gonadotropin-releasing hormone (GnRH) shown. (C) Proα1(I) mRNA stability. After incubation for 24 h with or without Ang II (1 μM), actinomycin D (10 μg/ml) was added to the medium and the incubation continued. At defined time points, proα1(I) mRNA levels were determined by Northern blot. Levels are expressed as a fraction of proα1(I) mRNA levels at time of actinomycin D application. Results shown are means ± SD from four separate experiments.