Fig. 6

EFTUD2 stabilizes c-MYC protein by inhibiting UPS-mediated degradation in CRC cells. A The top-ranked 3D docking conformation of EFTUD2 and c-MYC is shown using HDOCK. B Western blotting analysis detecting protein levels of c-MYC in SW480 and HCT116 treated with MG132 (10 µM) at different time intervals (0, 3, 6, 9 h) C Western blotting analysis showing changes in c-MYC expression before and after knockdown of EFTUD2 in SW480 and HCT116 simultaneously treated with MG132 (10 µM) for 6 h. D-G Protein half-life assay (D and E) and subsequent quantification (F and G) showing changes in c-MYC protein half-life upon modulation of EFTUD2 expression in SW480 and HCT116 treated with 20 µg/mL of CHX and transfected with sh-EFTUD2 or p-EFTUD2 plasmids. Protein is extracted at the indicated time points (0, 30, 60, 120 min). H and I Co-IP analysis detecting changes in the level of ubiquitination bound to c-MYC protein upon modulation of EFTUD2 expression in SW480 and HCT116 cells treated with MG132 (10 µM) and transfected with sh-EFTUD2 or Flag-EFTUD2 plasmids. J and K Co-IP analysis detecting changes in the level of c-MYC in SW480 and HCT116 cells transfected with a gradient of Flag-EFTUD2 plasmids. Each bar represents the mean values ± SD, **P < 0.01, ***P < 0.001