Fig. 2: CHFR is a ZEB1 E3 ligase.

a Transiently transfected SFB-ZEB1 and different concentrations of MYC-CHFR in HEK293T cells and immunoblotted with the MYC and FLAG antibodies. b In SUM159 cells, different concentrations of MYC-CHFR were transfected, followed by immunoblotting with the antibodies as indicated. c, d SFB-ZEB1 and MYC-CHFR were transiently transfected into HEK293T cells, and 50 μg ml⁻¹ CHX was added, and samples were collected at the indicated time points and analyzed by western blotting with FLAG and MYC antibodies. e, f In SUM159 stably expressed mock or CHFR cells, 50 μg ml⁻¹ CHX was added, and samples were collected at the indicated time points, followed by immunoblotting with antibodies against ZEB1, MYC, and β-actin. g LM2 was transduced by scramble or sh-CHFR cells, and samples were collected and immunoblotted by ZEB1, CHFR, and β-actin antibodies. h Transiently transferred SFB-ZEB1, MYC-CHFR, or MYC-CHFR∆RING into HEK293T cells. Immunoblotting of MYC and FLAG. i Transiently transfected MYC-CHFR or MYC-CHFR∆RING into SUM159 cells, followed by immunoblotting of ZEB1, MYC, and β-actin. j HEK293T cells were transiently transfected with SFB-ZEB1, MYC-CHFR, or MYC-CHFR ∆RING and added MG132 for 6 h before collecting samples, followed by pulling down with s-S protein beads and immunoblotting of MYC, HA, and FLAG.