Fig. 2

Normal lung fibroblasts were grown on tissue culture plastic in DMEM supplemented with 5% FBS to 60% confluence, serum deprived for 24 h in DMEM without serum and then treated with/without TGF-β1 at the indicated doses and durations. a and b Fas mRNA determined by quantitative real-time RT-PCR (qPCR); pooled data with an “n” of at least 7 independent replicates for each dose and time point. c and d Representative Western blots for Fas protein in whole cell lysates, with densitometric analysis. Densitometry represents data with at least 7 independent experimental replicates for each dose at the 24-h time point and 4 independent replicates at the 4- and 8-h time points. * p < 0.05 and ** p < 0.01 compared to untreated controls (One-way ANOVA with Tukey multiple comparison test). e and f Fas protein in whole cell lysates measured by ELISA. Data represent pooled results from at least 5 independent experimental replicates at the 24-h time points and 2–3 experimental replicates at the 4- and 8-h time points. ** p < 0.01 compared to control (One-way ANOVA with Tukey multiple comparison test). g Representative Western blot and (h) densitometry for Fas. n = at least 5 independent experimental replicates for each condition. *** p < 0.001 compared to untreated controls (One-way ANOVA with Tukey multiple comparison test)