Figure 3. SPOP, the adaptor protein of Cullin3 family E3 ligase, specifically interacts with Cdc20 through its N-terminal MATH domain.
A) Immunoblot (IB) analysis of whole cell lysates (WCL) and immunoprecipitates (IP) derived from 293T cells transfected with indicated constructs. 36 hours post-transfection, cells were pretreated with 10 µM MG132 for 10 hours before harvesting.
B) IB analysis of GST pull-down precipitates from 293T cell lysate with overexpression of HA-Cdc20 using bacterially purified GST and GST-SPOP proteins.
C) Schematic of SPOP with MATH and BTB domain.
D) IB analysis of WCL and anti-Flag IP derived from 293T cells transfected with indicated constructs. 36 hours post-transfection, cells were pretreated with 10 µM MG132 for 10 hours before harvesting.