Protein Quantification and Western Blot Analysis

Relative protein were extracted from tissue samples or cultured cells, and protein concentration was quantified by Lowry’s protein assay (Bio-Rad, Hercules, CA, USA) [19 (link)]. All protein samples were separated by 10% SDS-PAGE and then transferred onto polyvinylidene fluoride (PVDF) membranes.The primary antibodies used in this study were anti-BUB1B(#4116, Cell Signaling Technology, Beverly, MA, USA), anti-ZNF143(#PA5-72,658, Invitrogen, Waltham, MA, USA), anti-GLUT1(#ab115730, Abcam, Cambridge, UK), anti-LDHA(#ab101562, Abcam, Cambridge, UK), anti-PKM2 (#ab137852, Abcam, Cambridge, UK), anti-HK2 (#ab137852, Abcam, Cambridge, UK) and anti-Actin (#23,600-1-AP, Proteintech, China) antibodies. Western blot analysis was analyzed by Image J software.