FIGURE 4.

FBW7 dimerization is required for NFE2L3 degradation. A, NFE2L3 expressing construct was cotransfected with either HA-FBW7 or FLAG-FBW7ΔD constructs in HEK293T cells. These samples were analyzed by immunoblotting with antibodies specific for NFE2L3, HA, and FLAG. B, NFE2L3-expressing plasmid was cotransfected with either control vector, HA-FBW7, or FLAG-FBW7ΔD and HA-ubiquitin constructs in HEK293T cells and treated with MG-132 (10 μm) for 6 h before collection. NFE2L3 was immunoprecipitated from these lysates, and these samples were immunoblotted with NFE2L3 and HA antibodies to detect NFE2L3 ubiquitination.