Figure 3 Western blot analysis of autophagic and apoptotic markers in in MC3T3-E1 cells. (A) MC3T3-E1 cells were treated with vehicle control, 5-fluorouracil (5-FU) (20 µg/mL, 36 hours), rapamycin (500 ng/mL, 6 hours), 3-methyl adenine (3-MA) (10 nM, 12 hours), tumor necrosis factor-α (TNF-α) (20 ng/mL, 48 hours), rapamycin+TNF-α (rapamycin, 500 ng/mL, 6 hours; TNF-α, 20 ng/mL, 48 hours) or 3-MA+TNF-α (3-MA, 10 nM, 12 hours; TNF-α, 20 ng/mL, 48 hours). Expressions of LC3, p62 and cleaved caspase-3 were analyzed by western blot. β-Actin served as a loading control. (B) Quantitative data of western blot. Data were presented as mean±SD in triplicate. *p<0.05,** p<0.01 versus corresponding control.