Fig. 4

Degradation of Dvl2 in CYP2E1-overexpressing cells is ubiquitin–proteasome pathway dependent. A Control or CYP2E1-overexpressing HCC cells were transfected with empty vector or the Dvl2 expression vector. The protein level of Dvl2, β-catenin, and c-Myc were detected by western blot 48 h after transfection. B The OD450 value of MHCC-97H-CYP2E1, SMMC-7721-CYP2E1 and the respective control cells after transient transfections with empty vector or the Dvl2 expression vector. ^*P ≤ 0.05, ^**P ≤ 0.01, ***P ≤ 0.001 vs. CYP2E1 + vector. C The mRNA levels of Dvl2 and CTNNB1 in control and CYP2E1-overexpressing HCC cells were detected by qRT-PCR. D Co-immunoprecipitation analysis was applied to determine the ubiquitination level of Dvl2 in control and CYP2E1-overexpressing HCC cells. E Western blot analysis of Dvl2 protein levels in control and CYP2E1-overexpressing HCC cells at 1 h after treatment with the proteasome inhibitor MG132 (10 mM). F The protein level of KLHL12 in MHCC-97H-CYP2E1, SMMC-7721-CYP2E1 and the respective control cells was evaluated by western blot. G The interaction between Dvl2 and KLHL12 was determined by reciprocal co-immunoprecipitation analysis in control and CYP2E1-overexpressing HCC cells