Figure 7.

Opy2p is an integral membrane protein localized at the plasma membrane. (A) Plasma membrane localization of C-terminally GFP-tagged Opy2p in mid-log-phase end3Δ cells by fluorescent microscopy. (B) Subcellular localization of Opy2p. (Top) Cell extracts prepared from cells expressing GFP-tagged Opy2p were separated by centrifugation into pellet (P) and supernatant (S) fractions, with 15 and 100 indicating 15,000 × g and 100,000 × g, respectively. (Vector) A lysate containing no GFP-tagged Opy2p. (Bottom) Analysis of P15 was performed by treating the fraction with NaCl (0.5 M), Na₂CO₃ (200 mM at pH 11), urea (5 M), or SDS (5%) plus urea (5 M). Western blotting was performed using an antibody against GFP.