Fig 1: CD147 enhanced E‐cadherin ubiquitination and lysosomal degradation by Src activation. (A) Both HepG2 and HepG2‐CD147 cell lines were transfected with hemagglutinin‐ubiquitin plasmid, followed by detection of the interaction between E‐cadherin and Hakai by coimmunoprecipitation assay and the phosphorylation and ubiquitination of E‐cadherin by immunoprecipitation assay. An immunoglobulin G antibody was used as a negative control. (B) Western blotting detection of E‐cadherin expression and ubiquitination in HepG2‐CD147 and Huh‐7‐CD147 cells pretreated with MG132 (10 μM) or chloroquine (100 μM) for 12 hours. (C) Colocalization of E‐cadherin and lysosomes in the presence of chloroquine detected by immunofluorescence. Arrowheads indicate the colocalization of E‐cadherin with Lysosomes‐RFP. (D) Expression of Src phosphorylation (p‐Src) and Hakai was examined in CD147 overexpression, knockdown, and knockout cell lines by western blotting. (E) Effects of CD147 overexpression in the presence of Src inhibitor on E‐cadherin expression in total lysates and active β‐catenin in the nuclear fraction of cell lysates. (F) Both HepG2 and Huh‐7 cell lines were transfected with hemagglutinin‐ubiquitin plasmid, followed by detection of the interaction between E‐cadherin and Hakai by coimmunoprecipitation assay and the phosphorylation and ubiquitination of E‐cadherin under CD147 overexpression in the presence of Src inhibitor. An immunoglobulin G antibody was used as a negative control. Alpha‐tubulin, β‐actin, and lamin B were used as loading controls. Abbreviations: DAPI, 4′,6‐diamidino‐2‐phenylindole; IgG, immunoglobulin G; IP, immunoprecipitation.