Fig. 5

BMAL1 regulated carbon particles/PM2.5-induced autophagy via p53 protein. HBEs were incubated with control vector or BMAL1 expression vector for 72 h and treated with PM2.5 (200 μg/ml) or carbon particles (20 μg/ml) for 24 h prior to harvest of cell lysates, after which expression levels of p53 protein were measured by western blotting. A, B Representative images of Western blots of p53 were shown, and changes in relative density of p53 were presented. n = 6. C, D After transfection with human BMAL1 siRNA or negative control siRNA, HBEs were incubated with carbon particles or PM2.5 for 24 h, after which p53 protein were measured by western blotting and changes in relative density of p53 were presented. n = 6. E, F HBEs were incubated with control vector, BMAL1 expression vector or p53 expression vector for 72 h and treated with PM2.5 (200 μg/ml) or carbon particles (20 μg/ml) for 48 h prior to harvest of cell lysates, after which expression levels of p53, collagen-I, ATG5, LC3B-II, and p62 protein were measured by western blotting. Representative images of Western blots were shown, and changes in relative density of protein were presented. n = 3. Data are mean ± SEM of n individual experiments. *P < 0.05, **P < 0.01, ***P < 0.001 between indicated conditions (One-way ANOVA followed by the Bonferroni’s test)