Figure 3.

Amiodarone-induced autophagic machinery degrades p300 protein, activates FoxO1, and increases miR-449a expression. (A) SW480 cells were treated with amiodarone (10 μM) for 48 h. The levels of p300, AC-FKHR, FoxO1, and LC3 were measured using anti-p300, anti-AC-FKHR, anti-FoxO1, and anti-LC3 antibody by immunoblotting. β-actin was used as the internal control. (B) Cells were treated with amiodarone (10 μM) for 48 h. In the CQ group, the cells were treated with amiodarone (10 μM) together with CQ (50 μM) for 48 h. The p300 and LC3 levels were determined by immunoblotting using specific antibodies. β-actin was used as the internal control. (C) Colocalization of p300 and LC3 proteins were demonstrated by anti-p300 conjugated with rodamin (red) and anti-LC3 conjugated with FITC (green) antibody, respectively. The fluorescent change of the cells was investigated under a multi-photon confocal microscope. Quantification of colocalization was performed by counting 30 cells. The p-values were determined by Student’s t-test analysis.