X ray image film processor

Protein lysates were separated by SDS-PAGE in a 10% polyacrimide gel, then transferred to a polyvinylidene difluoride membrane (Millipore, USA). Membranes were incubated with antibody against human Bcl-w (Abcam; 1:200), Apaf-1 (Abcam; 1:500), cleaved Caspase-9 (Novus Bio; 1:1,000), cleaved Caspase-3 (CST; 1:1,000), Bax (Abcam; 1:2,000); or GAPDH (KangChen Bio-tech Inc., China). Peroxidase-conjugated goat anti-mouse IgG (H+L) (Southern Biotech) was used as the secondary antibody. The intensity of staining was visualized using an X-ray image film processor (Kodak, Japan).

The protein lysates were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in 10% polyacrylamide gel and then transferred to a polyvinylidene difluoride membrane (Merck Millipore, Billerica, MA, USA). The membrane was incubated with antibodies against human LC3B (ab192890, Abcam), Beclin-1 (#4122, Cell Signaling Technologies), MEK1/2 (ab178876, Abcam), phosphorylated (p)-MEK1/2 (ab194754, Abcam), Nrf-2 (#12721, Cell Signaling Technologies), and GAPDH (KC-5G4, Aksomics, Shanghai, China). Goat anti-mouse IgG (H + L) (KC-5G4, Southern Biotech, Birmingham, AL, USA) coupled with peroxidase was used as the second antibody. The intensity of staining was visualized using an X-ray image film processor (Kodak, Rochester, NY, USA).