BCNU increases TGFβ protein levels in 7WD10 cells. A) Cells were treated with BCNU at different concentrations as indicated and, after 48 hours, the conditioned media (CM) as well as lysates were immunoblotted to detect TGFβ protein. B) ImageJ quantitation revealed significantly increased TGFβ protein levels at 10.0 μM (73%, P < 0.01) and 20.0 μM (92%, P < 0.001) in the CM. Similarly in the lysates TGFβ levels were increased at the same concentrations of 10.0 μM (149%, P < 0.05) and 20.0 μM (154%, P < 0.01). For all samples, n = 4 ± SEM. *, P < 0.05, **, P < 0.01, ***, P < 0.001versus untreated controls by analysis of variance (ANOVA) followed by post-hoc test by Dunnett multiple comparisons. BCNU, 1, 3 bis (2-chloroethyl)-1-nitrosourea; SEM, standard error of the mean; TGFβ, transforming growth factor β.