Fig. 8
PFKL positively promotes YTHDF3 expression at protein level and PFKL interacts with YTHDF3 via EFTUD2 at the protein level. a PFKL and YTHDF3 protein expression by transiently transfecting small interfering RNA targeting PFKL mRNA into Sk-Hep-1, HepG2 and SNU449 cells for 12 hours. b. PFKL and YTHDF3 protein expression by transiently transfecting PFKL plasmids into Sk-Hep-1 and HepG2 cells for 12 hours. c-f. CCK8 proliferation assays, colony formation assays, transwell migration assays and transwell invasion assays after transiently transfecting small interfering RNA targeting PFKL into overexpression-YTHDF3 Sk-Hep-1, HepG2, SNU449 cells for 12 hours. The Image J software was used to quantified cells’ migration and invasion ability. Scale bar 200 μm. g. Endogenous co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and PFKL with Huh7 and SNU449 cells. h Exogenous co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and PFKL with YTHDF3-stably-overexpressed Sk-Hep-1 and HepG2 cells. i Co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and PFKL after co-transfecting YTHDF3 and PFKL plasmids (10 μg respectively) into 293 T cells for 48 h. j The processes of PFKL pre-mRNA splicing and YTHDF3 m6A modification to PFKL mRNA. k-m Co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and PFKL after transfecting EFTUD2 plasmids (10 μg) into Sk-Hep-1, Huh7 and SNU449 cells for 48 h. n Co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and EFTUD2 after co-transfecting YTHDF3 and EFTUD2 plasmids (10 μg respectively) into 293 T cells for 48 h. o Co-immunoprecipitation assays were performed to investigate the interaction between PFKL and EFTUD2 after co-transfecting PFKL and EFTUD2 plasmids (10 μg respectively) into 293 T cells for 48 h
p Co-immunoprecipitation assays were performed to investigate the interaction between YTHDF3 and PFKL after co-transfecting YTHDF3, PFKL and EFTUD2 plasmids (10 μg respectively) into 293 T cells for 48 h. Data are presented as mean ± SD (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001).