Figure 4.

Endometrial cancer-associated mutants of SPOP are defective in promoting ERα degradation and ubiquitination. (a) Distribution of the point mutations on the SPOP gene found in endometrial cancer samples. These mutations are exclusively located in the N-terminal MATH domain of SPOP. (b) Endometrial cancer-associated mutants of SPOP are defective in promoting ERα degradation. 293T cells were transfected with FH-ERα and wild-type or mutated SPOP constructs as indicated. After 24 h, cell lysates were prepared for WB analyzes. (c) Endometrial cancer-associated mutants of SPOP are defective in interacting with ERα. The 293T cells were transfected with the indicated constructs. After 24 h, cell lysates were prepared for co-IP assay with anti-FLAG antibody and WB analyzes. (d) Endometrial cancer-associated mutants of SPOP are defective in promoting ERα ubiquitination. The 293T cells were transfected with the indicated constructs. After 24 h, cells were treated with 20 μM MG132 for 4 h and cell lysates were prepared for immunoprecipitation and WB analyzes