Figure 1.

Cul2 neddylation status does not influence association with Rbx1. (A) ³⁵S-labeled in vitro-translated HA-Cul2(WT) or neddylation-defective HACul2(K689R) mutant was, where indicated, mixed with ³⁵S-labeled in vitro-translated myc-Rbx1 and immunoprecipitated with indicated antibodies. Bound proteins were resolved by SDS-PAGE and visualized by autoradiography. (B) HEK293A cells were transiently transfected with mammalian expression plasmids encoding myc-Rbx1, HA-Cul2(WT), or HA-Cul2(K689R), alone or in combination. Cells were lysed and immunoprecipitated with indicated antibodies. Bound proteins were resolved by SDS-PAGE and immunoblotted with anti-Cul2 (upper panel) and anti-myc (lower panel) antibodies. IP = immunoprecipitation.