Anti ras

Total cell or tissue extracts were extracted by cell lysis buffer followed by immunoblotting with anti-Ras (1:2,000; Proteintech), anti-β-actin (1:5,000; Proteintech), anti-ERK (1:1,000; Proteintech), anti-pERK (1:500; Proteintech), anti-MEK (1:1,000; Proteintech), and anti-pMEK (1:200; Proteintech). 25 µg of cell lysates was resolved with 12.5% SDS–PAGE and transferred to polyvinylidene fluoride. After 1-h blocking with blocking buffer, the membranes were incubated with primary antibody for 2 h at room temperature. After washing and 1-h incubation with secondary antibodies at room temperature, the blots were visualized using ECL.

METH was provided by Ningbo Public Security Bureau. N-Acetyl-L-cysteine (NAC) was purchased from Sigma-Aldrich. All drugs were dissolved in saline (0.9% NaCl) and administered to cells directly or via intraperitoneal injection. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and DMSO were purchased from MedChemExpress. The BrdU cell proliferation kit was purchased from BioVision. The primary and secondary antibodies for Western blotting, including anti-Ras, anti-β-actin, anti-ERK, anti-pERK, anti-MEK, and anti-pMEK, were purchased from Proteintech. Polyvinylidene fluoride, blocking buffer, and ECL were purchased from Millipore. The Active Ras Pull-Down and Detection Kit was purchased from Thermo Fisher Scientific.