FIGURE 6.

Bag6 binds gp78 and UbxD8 simultaneously. A, cells expressing Bag6, UbxD8 in the presence or absence of gp78-FLAG were lysed in the Nonidet P-40 lysis buffer. The lysates were first subject to immunoprecipitation with FLAG beads to pull down gp78-FLAG. The bound materials eluted with FLAG peptide were divided into two equal portions. One potion was incubated with control IgG whereas the other portion was incubated with anti-Bag6 antibodies to purify Bag6 and its associated proteins. The precipitated materials and a fraction of the inputs were analyzed by immunoblotting. B, diagram illustrating interactions between Bag6 and various ERAD factors. Solid arrows indicate direct interactions whereas dashed lines indicate association that may be mediated by an adaptor.