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. 2015 Dec 31;291(9):4487–4502. doi: 10.1074/jbc.M115.702514

FIGURE 2.

FIGURE 2.

OS9 interacts specifically with NKCC2 and the related NCC. A, co-immunoprecipitation of OS9 with NKCC2 and NCC in OKP cells. Cell lysates from cells transiently transfected with Myc-NKCC2 or Myc-NCC in the presence or absence of OS9-V5 were immunoprecipitated (IP) with anti-V5 antibody. Similar to NKCC2, NCC co-immunoprecipitated with OS9-V5 was detected by immunoblotting (IB) using anti-Myc (lanes 7 and 10). Again, the interaction with OS9 involves mainly the core glycosylated (immature) form of the co-transporters. 5% of total cell lysate (Lys) was resolved as a positive control. B, OS9 does not interact with ETB receptor. Cell lysates from OKP cells transiently transfected with the co-transporters proteins or ETB receptor in the presence or absence of OS9-V5 were immunoprecipitated with anti-V5 anti-body. Again, 5% of total cell lysate (Lys) was resolved as a positive control. In contrast to NKCC2 and NCC, ETB receptor protein was not recovered from OS9 immunoprecipitates (lane 10). IgGH, the heavy chain of IgG.