Fig. 9.

Disruption of NKCC1 COOH-terminal domain disrupts the cotransporter interaction with cell trafficking and polarity machinery. Single-cell suspension of MDCK stably transfected with EGFP-NKCC1-WT (A), EGFP-NKCC1-DFX (B), EGFP-NKCC1–241 (C), EGFP-NKCC1–242 (D), and EGFP-NKCC1–244 (E) were grown on 3D in Matrigel for 6–48 h. We then followed these five MDCK clones overtime (F–T). After each time point, cells were fixed and immunostained with anti-Gp135/podocalyxin. Bar = 10 μm WT, wild-type; DFX, Δ187aa, 241, LL1031-1032AA; 242, LL1192-1193AA; 244, LL1031-1032AA + LL1192-1193AA. Bars = 10 μm. GFP, green fluorescent protein; MDCK, Madin-Darby canine kidney; NKCC1, Na⁺-K⁺-2Cl⁻ cotransporter-1. Data are representative of 3 individual experiments where 5 cysts per experiment were imaged (Total, n = 15).