ACA was dissolved in dimethyl sulfoxide (DMSO; Fisher Scientific, USA) to serve as a stock solution. The autophagy inhibitors used were 3-methyladenine (3-MA; Calbiochem, USA) and chloroquine diphosphate (CQ; Sigma, USA). The 3-MA was dissolved directly in culture media and the stock solution CQ was prepared in distilled water. Acridine orange solution was obtained from Sigma. Roswell Park Memorial Institute-1640 (RPMI-1640) medium was purchased from GE Healthcare HyClone (USA) whereas alpha-Minimum Essential Medium (α-MEM) was obtained from Nacalai Tesque (Japan). Fetal bovine serum (FBS) was from Sigma (USA).
Primary antibodies included rabbit antibodies against microtubule associated protein 1 light chain 3-I/II (LC3-I/II), p62, GAPDH and mouse antibodies against Beclin-1 were used. Anti-rabbit IgG and anti-mouse IgG horseradish peroxidase (HRP)-conjugated secondary antibody were used. All the antibodies were purchased from Cell Signaling Technology (USA).
Primary antibodies included rabbit antibodies against microtubule associated protein 1 light chain 3-I/II (LC3-I/II), p62, GAPDH and mouse antibodies against Beclin-1 were used. Anti-rabbit IgG and anti-mouse IgG horseradish peroxidase (HRP)-conjugated secondary antibody were used. All the antibodies were purchased from Cell Signaling Technology (USA).