Fig. 9From: A functional loop between YTH domain family protein YTHDF3 mediated m6A modification and phosphofructokinase PFKL in glycolysis of hepatocellular carcinomaPFKL protein positively regulates YTHDF3 protein expression via inhibiting ubiquitination of YTHDF3 protein by EFTUD2. a Confocal assays of three group with Sk-Hep-1 cells incubated PFKL and EFTUd2 antibodies, YTHDF3 and EFTUd2 antibodies, or YTHDF3 and PFKL antibodies to investigate the localization between YTHDF3, PFKL and EFTUD2 (upper). Image J software was used to qualified the green, red and yellow fluorescence colocalization signal intensity, and calculated relative Pearson’s and Mander’s coefficients (lower). Scale bar 100 μm. b Multi-label immunofluorescence staining of different HCC patients to investigate the localization between YTHDF3, PFKL and EFTUD2 (upper). Image J software was used to qualified the green, red and yellow fluorescence colocalization signal intensity (lower). Scale bar 100 μm. c Ubiquitination assays of SNU449 cells co-transfected ubiquitin, YTHDF3 and PFKL plasmids. 20 nM MG-132 was added to RPMI1640 culture medium and cells were incubated continuously for 8 hours. d. Ubiquitination assays of SNU449 cells co-transfected ubiquitin, YTHDF3 and EFTUD2 plasmids. 20 nM MG-132 was added to RPMI1640 culture medium and cells were incubated continuously for 8 hours. e Ubiquitination assays of SNU449 cells co-transfected ubiquitin, YTHDF3, PFKL and EFTUD2 plasmids. 20 nM MG-132 was added to RPMI1640 culture medium and cells were incubated continuously for 8 hours. f Molecular mechanism of the relationship between YTHDF3, PFKL and EFTUD2 in HCC progressionBack to article page