Anti glut1 antibody

Trametinib and U0126 were purchased from MedChemExpress (Monmouth Junction, NJ, United States). Dimethyl sulfoxide was used to dissolve Trametinib powder to an initial concentration of 10 mM and preserved in −80°C. The CCK-8 kit was obtained from Dojindo (Rockville, MD, United States). The Annexin V-FITC/PI apoptosis detection kit and Annexin V-PE/7-ADD apoptosis detection kit were purchased from Nanjing Vazyme Biotech (Nanjing, China). Penicillin and streptomycin were obtained from Gibco (Thermo Fisher Scientific, Waltham, MA, United States). DMEM were purchased from CORNING (Corning Life Sciences, Bedford, MA, Unied States). The fetal bovine serum (FBS) was purchased from TBD (Tianjin, China). Lipofectamine 3,000 and TRIzol™ Reagent were obtained from Life Technologies (Carlsbad, CA, United States). XF96 seahorse assay plate was purchased from Agilent (Santa Clara, CA, United States). The antibodies used were as follows: anti-PKM2 antibody (#15822-AP), anti-c-myc antibody (#67447-1-lg), anti-GLUT1 antibody (#21829-1-AP), anti-LDHA antibody (#19987-1-AP), anti-β-actin antibody (#66009-1-lg), MEK1/2 polyclonal antibody (#11049-1-AP), ERK1/2 polyclonal antibody (#16443-1-AP), and anti-PCNA antibody (10205-2-AP) from Proteintech (Wuhan, China); and anti-Ki67 antibody (ab279653) from Abcam (Cambridge, United Kingdom).

Western blot was performed to measure protein expression levels of HIF-1α, PDK1, GLUT1, HK2, Ubiquitin, UFL1, and BRE1B. And following antibodies were used: anti- HIF-1α antibody (1:300; Proteintech, United States), anti-GLUT1 antibody (1:1,000; Proteintech, United States), anti-HK2 antibody (1:2000; Cell Proteintech, United States), anti-PDK1 antibody (1:2000; Proteintech, United States), anti-ubiquitin antibody (1:1,000; Proteintech, United States), anti-UFL1 antibody (1:1,000; Bethl Laboratories, United States) and anti- BRE1B antibody (1:1,000; Bethl Laboratories, United States).