FIGURE 3.

Wsc1* requires Rsp5p and Doa4p for entry into the MVB pathway. A, wild type, Δpep4, rsp5-1, and Δpep4 rsp5-1 cells expressing Wsc1* were grown to log phase and shifted to 37 °C for 1 h. Cell preparation and image acquisition were performed as described in the legend to Fig. 1B. Scale bar, 5 μm. B, indirect immunofluorescence of wild type, Δpep4, Δdoa4, and Δpep4Δdoa4 strains expressing Wsc1*. Scale bar, 5 μm. C, Western blot analysis of wild type, Δpep4, rsp5-1, and Δpep4 rsp5-1 cells expressing Wsc1* after a 1-h shift to 37 °C. D, Wsc1* expression in wild type, Δpep4, Δdoa4, and Δpep4Δdoa4 cells was analyzed by immunoblotting as described in the legend to Fig. 2C.