Quantitative Protein and Gene Expression

Western blotting and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) were used for molecules detection and performed as previously described (Wu et al. 2015 (link)). Antibodies against HK2 (22029-1-AP,), AKT1 (10176-2-AP), LDHA (19987-1-AP, ), GLUT1 (21829-1-AP), and GSK3β (22104-1-AP) all purchased from Proteintech; β-Actin (#4970), phosphor-AKT1 (Ser473, #4060), and phosphor-GSK3β (Ser9, #5558), all purchased from Cell Signaling Technology, were used to assess the expression of an individual protein. Secondary antibodies were used at 1:5000 dilutions. The primer sequences for RT-qPCR involved in this study were shown in Supplemental Table 2.