FIGURE 7.

FBW7-mediated degradation of NFE2L3 is GSK3-dependent. A, NFE2L3 expression plasmid was cotransfected with either HA-FBW7 alone or in combination with shRNA targeting GSK3B in HEK293T cells. Lysates were analyzed by immunoblot with antibodies specific for NFE2L3 and HA antibody to detect FBW7 as well as GSK3. B, plasmid coding for NFE2L3 was cotransfected with HA-ubiquitin construct and GSK3B shRNA or in combination with FBW7 shRNA in HEK293T cells and treated with MG-132 (10 μm) for 6 h before collection. NFE2L3 was immunoprecipitated, and these samples were immunoblot-analyzed with antibodies specific for NFE2L3, HA, and GSK3.