Fig. 8.

a to d, representative pictures of in situ TUNEL staining in tibial bone marrow taken from 36% EtOH-treated mice (a) and corresponding PF controls (b), 30% EtOH-treated mice (c), and 30% EtOH/VitD-treated mice (d) as described under Materials and Methods. Arrows indicate apoptotic cells. e, EtOH exposure (0–50 mM) increased cleaved caspase-3 expression in differentiating osteoblastic cells in a dose-dependent manner. Pretreatment with 1,25(OH)₂D₃ protected against EtOH-induced apoptosis in osteoblastic cells treated with EtOH. Apoptosis was assessed by Western blot with caspase-3 expression quantified by densitometry. Band densities were corrected for total protein loaded by staining with 0.1% amido black. Bars indicate mean ± S.E.M. of triplicate determinations. Statistical differences were determined by one-way ANOVA followed by Student-Keuls post hoc analysis. P < 0.05 for EtOH treatment a < b < c.