Fig. 5.

Effect of WNK463 on physiological stimuli of KCC in red cells from patients with SCA. Red cells (20% Hct) were pre-incubated in N-MBS in the presence or absence of WNK463 (40 nM) at 37 °C in air. They were then equilibrated in Eschweiler tonometers for 20 min at 150 mmHg, after which aliquots were diluted tenfold into flux tubes containing buffers varying in pH, osmolarity, or urea content, all in the continued presence or absence of WNK463, and KCC activity was measured as described in the legend to Fig. 1. a Effect of WNK463 on pH-dependent KCC activity, n = 6. b Effect of WNK463 on KCC activity in 10% shrunken (SH) or 10% swollen (SW) red cells, n = 5. c Effect of WNK463 on urea-induced KCC activity, n = 6. d Impact of WNK463 on pH, volume, and urea-stimulated KCC activity. Histograms represent means ± SEM of n individual samples. * p < 0.05, ** p < 0.01, *** p < 0.001 compared to indicated condition, ^# p < 0.05 compared to fold change in absence of WNK463