Research Paper Volume 13, Issue 7 pp 10603—10618

Tumor suppressor DCAF15 inhibits epithelial-mesenchymal transition by targeting ZEB1 for proteasomal degradation in hepatocellular carcinoma

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Figure 2. DCAF15 binds with ZEB1 protein. (AB) Ectopically expressed DCAF15 and ZEB1 can bind with each other. (A) Flag-DCAF15 and Myc-ZEB1 were co-expressed in HEK293T cells, and immunoprecipitated with FLAG-M2 agarose beads. (B) Similar Co-IP assay was performed between Flag-ZEB1 and Myc-DCAF15. (CD) Endogenous interaction between DCAF15 and ZEB1. (C) SMMC-7721 cells were treated with 10μM MG132 for 6h. Cell lysates were precipitated using anti-ZEB1 antibody or with IgG (mock IP) and coprecipitated DCAF15 was detected by western blotting. (D) SMMC-7721 cells were treated with 10 μM MG132 for 6h. Cell lysates were precipitated using anti-DCAF15 antibody or with IgG (mock IP) and coprecipitated ZEB1 was detected by western blotting. (E) Flag-DCAF15 and Myc-ZEB1 were co-expressed in Hep3B and SMMC-7721 cells respectively and visualized by fluorescence microscopy. Scale bars, 30 μm.