Primary lung fibroblasts (PLFs) were isolated from the lungs of C57BL/6 mice, and after 3-7 passages, cells were cultured ± alcohol (60 mM) for up to 72 hours, messenger RNA was isolated at 24, 48, and 72 hours and total protein was isolated at 72 hours. (A) At 24 hours of treatment, PLFs showed a trend toward an increase in Smad7 gene expression; however, gene expression was reduced by approximately 30% at 48 hours and nearly eliminated at 72 hours (as reflected by Smad7 mRNA levels, normalized to 18s). (B) There was an approximate 30% reduction of Smad7 protein expression at 72 hours as determined by Western immunoblot analyses (normalized to GAPDH; representative gels shown above summary data). *P<0.05 compared to untreated groups.