Evaluation of autophagy flux
Cardiomyocytes were seeded in 35 mm dishes at a density of 1.5 106 cells/well in medium containing 2% FBS. To evaluate autophagy flux, the cells were incubated with Doxo 1 uM and Baf A 10 uM for 24 h. Cell lysates were analyzed by Western blotting with anti-LC3 and GAPDH antibodies. The autophagic flux was expressed in relation to untreated cultures with Baf A.