Fig. 8From: A Christianson syndrome-linked deletion mutation (∆287ES288) in SLC9A6 disrupts recycling endosomal function and elicits neurodegeneration and cell deathExpression of NHE6 does not affect trafficking of the EGF receptor in HeLa cells. a HeLa cells were transiently transfected with NHE6v1GFP WT (upper panels) or ΔES (lower panels). Forty-eight h after transfection, cells were loaded with Alexa Fluor647-labelled EGF (EGF-AF647, 100 ng/mL) for 5 min, fixed in 4 % paraformaldehyde, permeabilized, mounted onto glass slides and then examined by confocal microscopy. Footprints of the transfected cells within the field of view are indicated as white dotted outlines and were derived from the transmitted light images (far left panels). Scale bars represent 10 μm. b Uptake of EGF-AF647 (100 ng/ml, 5 min) in 104 GFP-positive HeLa cells expressing GFP, NHE6v1GFP WT or ΔES measured by flow cytometry. Data were normalized and represent mean ± S.E.M. of five different experiments. c Uptake of EGF-AF647 (100 ng/ml, 5 min) in HeLa cells expressing GFP, NHE6v1GFP WT or ΔES in the absence (−) or presence (+) of 10-fold excess unlabeled EGF (1 μg/ml). Median fluorescence intensity (M.I.F.) of EGF-AF647 was measured in 104 GFP-positive cells by flow cytometry and values represent the mean ± S.E.M. of three different experiments. Significance was established using a one sample Student t-test, **p < 0.001Back to article page