Wl02891

Monoclonal anti-Actin Beta (ACTB) antibody (AC026, ABclonal Technology, Wuhan, China), monoclonal anti-GFP antibody (AE012, ABclonal Technology), monoclonal anti-RFP antibody (AE020, ABclonal Technology), monoclonal anti-His antibody (AE003, ABclonal Technology), polyclonal anti-phospho-AKT antibodies (AP0140, ABclonal Technology), polyclonal anti-phospho-FOXO antibodies (AP0684, ABclonal Technology), mouse control IgG (AC011, ABclonal Technology), rabbit control IgG (AC005, ABclonal Technology), Horseradish Peroxidase (HRP)-conjugated goat anti-rabbit IgG light chain (AS061, ABclonal Technology), polyclonal anti-AKT antibodies (WL0003b, Wanleibio, Shenyang, China), polyclonal anti-FOXO antibodies (WL02891, Wanleibio), Alkaline phosphatase (AP)-conjugated horse anti-mouse IgG (ZB2310, ZSGB-BIO, Beijing, China), and AP-conjugated goat anti-rabbit IgG (ZB2308, ZSGB-BIO).

After total proteins were harvested, a western blot was performed according to standard methods to detect protein expression. Briefly, total proteins were separated by SDS-PAGE (P1200, Solarbio, China) and then transferred to polyvinylidene difluoride membranes (1620184, Bio-Rad, USA). After blocking, the membranes were first imprinted with YB-1 (ab12148, Abcam, UK), P53(WL01919, Wanleibio, China), P21(WL0362, Wanleibio, China), FOXO₃ (WL02891, Wanleibio, China) or GAPDH (ab9485, Abcam, UK) primary antibodies for 12 h at 4 °C followed by membrane wash and secondary antibodies (#14708, Cell Signaling Technology, USA) incubation for 1.5 h. The protein band signals were detected by ECL chemiluminescent substrate (E412-01, Vazyme, China) in ChemiDoc™ XRS + with Image Lab™ software (Bio-Rad, USA).