Figure 5.
Wsc1-L63R lacks an ERAD determinant in its luminal domain. (A) A schematic diagram of Wsc1-L63R and Wsc1-L63RLuminal. Asterisk indicates the L63R mutation. S.S., signal sequence; TM, transmembrane domain; Cyt, cytoplasmic domain. (B) Wild-type cells expressing Wsc1-L63RLuminal were processed as in Figure 3D. The cells were stained with anti-HA and anti-Kar2p primary antibodies followed by Alexa Fluor 488 goat anti-mouse and Alexa Fluor 594 goat anti-rabbit secondary antibodies. Cells were visualized by confocal microscopy. Scale bar, 5 μm. (C) The turnover of Wsc1-L63R and Wsc1-L63RLuminal in the wild-type strain was measured by pulse-chase analysis as described in Figure 2A. (D) Pulse-chase analysis was performed in wild-type and sec12-4 cells expressing Wsc1-L63RLuminal as described in Figure 1B.