Table 1. Quantification of total and glycosylated sub-fraction of KCC2 at the cell surface and in whole cell lysate.

	Glycosylated plus unglycosylated KCC2 at the surface (fraction of total, % of wild-type)	Glycosylated KCC2 (% of glycoslyated plus unglycosylated)
		Surface fraction	Total fraction
WT	100±±0%	50±13%	43±1%
L311H	34±7%**	22±5%*	17±1%**
G551D	41±10%*	32±7%	38±1%*
L426P	55±13%*	10±3%**	16±4%*

WT, wild-type.

n=4. *P<0.05, **P<0.005 (compared with WT, one-sample Student's t-test).

All three mutants showed significantly reduced levels of cell-surface over total expression of KCC2 when compared with WT with reduced levels of glycosylated KCC2 protein evident at the cell surface and also in total cell lysates (Figure 4).