FIGURE 1.

Identification of two additional trafficking signals in NKCC2 COOH-terminal domain. A, predicted topology of NKCC2. The distal region of NKCC2 COOH terminus, downstream from Y998X mutation (indicated by arrow) is enriched of di-leucine like motifs); ¹⁰³⁸LL¹⁰³⁹, ¹⁰⁴⁸LI¹⁰⁴⁹, ¹⁰⁵⁰VL¹⁰⁵¹, ¹⁰⁶⁴LL¹⁰⁶⁵, and ¹⁰⁷²IL¹⁰⁷³. B, immunoblot analysis of OKP cells expressing wild-type or mutant NKCC2 proteins as indicated. Cells were transiently transfected with WT NKCC2 or mutated NKCC2 proteins; ¹⁰³⁸AA¹⁰³⁹, ¹⁰⁴⁸AA¹⁰⁴⁹, ¹⁰⁵⁰AA¹⁰⁵¹, ¹⁰⁶⁴AA¹⁰⁶⁵, and ¹⁰⁷²AA¹⁰⁷³. Forty-eight hours post-transfection, total cell lysates were separated by SDS-PAGE and probed by anti-Myc antibodies. The positions of the core (immature) and complex-glycosylated (mature) proteins are indicated. C, immunoblot analysis of HEK cells expressing wild-type or mutant NKCC2 proteins, as indicated, using anti-Myc antibodies.