Fig. 6.

ENO2 regulates CCND1-mediated cell cycle progression via PKM2 in HNSCC cells. a, b Effect of PKM2 expression restoration on cell cycle in ENO2 knockdown UM-1 and Cal27 cells. c, d The nuclear translocation of PKM2 in ENO2 knockdown and control UM-1 and Cal27 cells. Representative confocal images and quantitative data from 100 cells are shown in (c) and (d), respectively. e Effect of ENO2 knockdown on nuclear and cytoplasmic PKM2 protein levels in UM-1 and Cal27 cells. f Effect of PKM2 expression restoration on CCND1 expression in ENO2 knockdown UM-1 and Cal27 cells. g Effect of compound 3k (Com 3k) on CCND1 expression in ENO2 overexpressing (ENO2 O/E) and control Cal33 and HSC3 cells. h Effect of AZD5363 on nuclear PKM2 protein levels in ENO2 overexpressing and control Cal33 and HSC3 cells. i Effect of U0126 on nuclear PKM2 protein levels in ENO2 overexpressing and control Cal33 and HSC3 cells. In (h-i), the purity of the nuclear fractions was indicated by the nuclear PCNA protein and the absence of cytosolic GAPDH protein. j The binding of PKM2 protein on the CCND1 gene promoter in ENO2 knockdown and control UM-1 and Cal27 cells determined by ChIP-qPCR assays. **p<0.01