Antibody against gapdh

GRS was a mixture of ginsenoside Rb1, ruscogenin and schisandrin (6:0.75:6). Schisandrin and ginsenoside Rb1 were obtained from Zelang Bio-Technology Co., Ltd (Nanjing, China). Ruscogenin was seperated in the laboratory and the purity was higher than 99%. Fetal bovine serum (FBS) was received from ScienCell (Carlsbad, CA, USA). Dulbecco’s modified Eagle medium (DMEM) was purchased from GIBCO/BRL (Life Technologies, Carlsbad, CA, USA). N-acetyl cysteine (NAC) was purchased from Sigma-Aldrich (St. Louis, MO, USA). 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) was purchased from AMRESCO (Cleveland, OH, USA). The kits for determination of lactate dehydrogenase (LDH), the malondialdehyde (MDA) and the fluorescent kit for 2′, 7′-dichlorofluorescein diacetate (DCFH-DA) were obtained from Beyotime Institute of Biotechnology (Shanghai, China). Protease inhibitor, RIPA lysis buffer, and enhanced chemiluminescene (ECL) reagent were obtained from Vazyme Biotech (Nanjing, China). Antibody against GAPDH was purchased from Bioworld Technology (St. Louis Park, MN, USA), antibody against JNK, p-JNK, p38, p-p38, ERK1/2, p-ERK1/2 and Jak2, p-Jak2, Stat3, p-Stat3 were obtained from Cell Signaling Technology (Boston, MA, USA).

Human CRC cells SW1116 were cultured in RPMI 1640 medium, and HEK293T cells were cultured in DMEM medium supplemented with 10% fetal bovine serum, 100 units of penicillin/ml and 100 ng of streptomycin/ml. Cells were incubated at 37°C in a humidified chamber supplemented with 5% CO₂. Antibodies against KSR1, p-ERK1/2 and ERK1/2 were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibody against GAPDH was from Bioworld Technology (Atlanta, Georgia 30305, USA). The growth factor reduced Matrigel was from BD Biosciences (Bedford, MA, USA).

The RPMI-1640 medium, DMEM-F12 medium and heat-inactivated fetal bovine serum (FBS) were from GIBCO (Gaithersburg, MD, USA). The RNAiso Plus was purchased from Takara (Shiga, Japan). TransScript First-Strand cDNA Synthesis SuperMix and TransStart Top Green qPCR SuperMix were purchased from TransGen (Beijing, China). PKM2 antibody was obtained from Cell Signaling Technology (Danvers, MA, USA). Antibodies for p53, Drp1 and ETC complexes were obtained from Proteintech (Chicago, USA). Antibody against GAPDH was from Bioworld Technology (Minneapolis, MN). Actinomycin D (RNA synthesis inhibitor), Cycloheximide (CHX, protein synthesis inhibitor) and MG-132 (the proteasome inhibitor) were obtained from Sigma (St. Louis, USA).

Human embryonic kidney cell line 293 T (HEK293T), Breast cancer cell lines (MCF-7, MDA-MB-231), and mammary epithelia cell (MCF10A), were obtained from American Type Culture Collection. Cells were incubated at 37 °C in a humidified chamber supplemented with 5% CO2. Antibodies against IRS1, p-AKT, AKT, p-ERK1/2 and ERK1/2 were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibody against GAPDH was from Bioworld Technology (Atlanta, Georgia 30305, USA). The growth factor reduced Matrigel was from BD Biosciences (Bedford, MA, USA).