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. 2005 Sep 26;102(40):14132–14138. doi: 10.1073/pnas.0505006102

Fig. 1.

Fig. 1.

Interaction of E3 ligases with components of the Derlin-1 complex. (a) Extracts from 293T cells, transfected with an empty vector (control) or with constructs encoding Myc-tagged wild-type Hrd1 (Hrd1 wt Myc), or Myc-tagged Hrd1 mutant containing a mutation in its ring finger domain (Hrd1 C291A Myc), were analyzed by immunoprecipitation (IP) with Myc antibodies, followed by immunoblotting (IB) with the indicated antibodies. Ten percent of the extracts were analyzed directly by immunoblotting (input). (b) A detergent extract of dog pancreatic microsomes was analyzed by immunoprecipitation (IP) with the indicated antibodies, followed by immunoblotting (IB) with various antibodies. The asterisks indicate proteins crossreacting with the antibodies. (c) As in a, except that cells expressed Hrd1-Myc, FLAG-tagged gp78 (gp78-FLAG), or both. The asterisk indicates residual FLAG-gp78 left after incomplete stripping of the FLAG immunoblot (Top). (d) As in a, except that cells expressed FLAG-tagged Parkin (FLAG-Parkin). IgG, immunoglobins detected by the secondary antibodies.