Figure 1.

ENaC is ER localized in yeast. (A) Schematic representation of the ENaC subunits demonstrating topology and N-linked glycosylation sites (triangles), which initially reside in the ER lumen. (B) Cell lysates from wild-type yeast expressing C-terminally HA-tagged α-, β-, or γ-ENaC treated with endoglycosidase H (Endo H). A small amount of glycosylated β-ENaC was evident due to incomplete conversion. In all cases, the fastest migrating species corresponds to the predicted size for each unglycosylated subunit. (C) Localization of ENaC by immunofluorescence. Panels left to right show the same fluorescent field for DAPI nuclear staining, Kar2 (yeast BiP), used as an ER marker, and HA antibody for ENaC, respectively.