A HEK293 cells were nucleofected with the indicated WT, R952H, and R1049C KCC2 constructs. After 3 days, cells were lysed, and Western blotting was performed using anti-phospho-S940 KCC2 and total anti-KCC2 antibodies, as described in Materials and Methods. A representative Western blot is shown.
B Quantification of total phosphorylation of S940 relative to total KCC2 expression for each construct. 4 separate nucleofections were performed. *P < 0.05, one-way ANOVA test; All pairwise multiple comparison procedures (Tukey’s test). Error bars represent the mean ± s.e.m.