Figure 4.

Effects of cycloheximide (CHX, 30 μM, left-hand ordinate) or TGF-β in absence or presence of 30 μM CHX (right-hand ordinate) on expression of prepro-ET-1 (ppET-1) in MRC-5 human lung fibroblasts. After dissemination, cells were cultured for 24 h in presence of 10% FCS followed by 1.5 or 4.5 h in presence of 1% FCS and absence or presence of cycloheximide (30 μM, left-hand ordinate) or 1 h TGF-β (1 ng·mL⁻¹) and/or cycloheximide (30 μM, 1.5 h, i.e. present 30 min before TGF-β). Thereafter, total RNA was isolated, treated with DNase and used for quantitative real time PCR. Height of columns: ppET-1 mRNA (ΔCt over GAPDH) is expressed as % of the control of the individual cell preparation, given are means ± SEM of n ≥ 6. Significance of differences: ***P < 0.001 versus respective Ctr.; ⁺⁺⁺P < 0.001 versus respective value in presence of TGF-β or CHX alone.