Fig. 12

Expression of NHE6∆ES causes a reduction in transferrin uptake and colocalization in primary hippocampal neurons. a Confocal images of the cell bodies of primary hippocampal neurons transfected with GFP (green) and NHE6_ChFP WT or ∆ES (red) and incubated with Alexa Fluor⁶³³-tagged transferrin (Tf-AF⁶³³) (pseudo-coloured blue) to assess endocytotic transferrin uptake and colocalization. Images show each channel individually, with merged images of the NHE6_ChFP and Tf-AF⁶³³ channels. b Quantitative summary of mean ± S.E.M. thresholded Mander’s coefficients, a measure of colocalization, between Tf-AF⁶³³ and NHE6_ChFP from four experiments. By the Mander’s coefficient, the majority of Tf-AF⁶³³ was colocalized with NHE6_ChFP WT than the inverse. The degree of colocalization was decreased with the NHE6_ChFP ∆ES mutant. *: p < 0.0001; **: p < 0.05, independent Student’s t-test, two-tailed. Scale bar: 10 μm