Figure 2.
(a) Immunoblot of immunoprecipitated oocyte membranes probed with anti-NCC antibody. Oocytes injected with NCC, NCC + WNK4, and NCC + WNK4 + WNK1 express the same amount of core-glycosylated protein. The ratio of mature glycosylated to core-glycosylated NCC is the same in all three groups. (b) Immunocytochemistry of oocytes stained with anti-NCC. Representative oocytes injected with NCC, NCC + WNK4, and NCC + WNK4 + WNK1 are shown. (c) Immunoblot of surface-biotinylated NCC. Only the mature glycosylated form of NCC was evident in this blot (data not shown). Coexpression of WNK4 with NCC reduced surface NCC expression by 85% (see Results). Coexpression with WNK1 and WNK4 restored surface NCC levels toward levels observed with NCC alone.