Anti mtor

Following the previously established procedure^{24 (link)}, post 24 h naringenin treatment, the cells were lysed using RIPA buffer. Protein expressions of β-actin, MMP9, PI3K, AKT, and Caspase-3 were assessed through SDS-PAGE, transferred onto a PVDF membrane, the blots were cut prior to hybridization with antibodies, and cropped blots incubated overnight at 4 °C with primary antibodies including anti-MMP9 (Beyotime, 1:1000), anti-mTOR (Beyotime, 1:1000), anti-PI3K (Cell signaling, 1:1000), anti-p-PI3K (Cell signaling, 1:1000), anti-AKT (Cell signaling, 1:1000), anti-p-AKT (Cell signaling, 1:1000), anti-EGFR (ABclonal, 1:1000), anti-Caspase-3 (Beyotime, 1:1000), and anti-β-actin (Cell signaling, 1:1000). Secondary antibodies used were either GAM (goat anti-mouse IgG) or GAR (goat anti-rabbit IgG) (ABclonal, 1:2000).

The tissue microarray was purchased from Alenabio Co. Ltd. Foetal bovine serum (FBS) and Dulbecco's Modified Eagle's Medium (DMEM) were obtained from Thermo Fisher Inc. Anti‐RPL22L1 and the secondary antibodies were purchased from Proteintech. CCK‐8 kit, transwell chambers, enhanced chemiluminescence reaction kit (ECL), bovine serum albumin (BSA), LY294002, anti‐PI3K (p85), anti‐p‐PI3K (p85), anti‐Akt, anti‐p‐Akt (Ser473), anti‐mTOR, anti‐p‐mTOR (Ser2448) and anti‐β‐actin were obtained from Beyotime Biotechnology. RPL22L1 lentiviral activation particles and RPL22L1 shRNA lentiviral particles were purchased from Santa Cruz.