FIGURE 6:

Yos9p is required for targeting of CPY*-CPY to Hrd1p. (A) Cell lysates were prepared from wild-type or yos9Δ cells transformed with a vector alone, a vector expressing CPY*-CPY, or a vector expressing CPY*(N479Q)-CPY and subjected to immunoprecipitation with anti-FLAG agarose. Cell lysates (Input) and immunoprecipitated materials (IP: FLAG) were analyzed by SDS–PAGE and immunoblotting with anti-CPY (IB: αCPY), anti-Hrd3p (IB: αHrd3p), and anti-Hrd1p (IB: αHrd1p) antibodies. Relative amounts of Hrd3p or Hrd1p coprecipitated with the fusion proteins were quantified. Error bars represent SDs from three independent experiments. The amount of Hrd3p or Hrd1p coprecipitated with CPY*-CPY in wild-type lysate was set to 100%. (B) Cell lysates were prepared from wild-type cells transformed with a vector alone or wild-type, hrd1Δ, or hrd3Δ cells transformed with a vector expressing CPY*-CPY and subjected to immunoprecipitation as in Figure 3B.