Figure 5. Effects of Hmgb1 knockdown and overexpression on SNCA-induced autophagy impairment. (A) iPC12 cells were transfected with Hmgb1 siRNA or control siRNA for 48 h and then induced to express SNCA for another 24 h. The expression of SNCA, HMGB1 and LC3 were determined by western blot. Experiments were performed 3 times with similar results and the representative blots were shown. (B) Relative intensity of LC3-II is normalized to the corresponding ACTB levels. Data are presented as the mean ± SD from 3 independent experiments. *P < 0.05 vs. uninduced and untransfected (UT) controls. (C) iPC12 cells were treated with 2 μg/ml Dox and transfected with Flag-HMGB1 plasmid or the relative empty vector for 24 h. The expressions of LC3-II and HA-SNCA were determined by western blotting. Experiments were performed 3 times with similar results and the representative blots were shown. (D) Relative levels of LC3-II are normalized to those of ACTB. Data are presented as the mean ± SD from 3 independent experiments. ^# P < 0.05 vs. uninduced and untransfected (UT) controls; *P < 0.05 vs. induced and N.T. controls.