The location of NBCe1‐A‐EGFP is disclosed using an anti‐EGFP primary antibody followed by an Alexa488‐conjugated secondary antibody. The distribution of the basolateral marker Na,K‐ATPase is disclosed using an anti‐Na,K‐ATPase primary antibody followed by an Alexa594‐conjugated secondary antibody. A, cells transfected with WT‐EGFP exhibit lateral distribution of NBCe1‐A‐EGFP. B, cells transfected with R510H‐EGFP exhibit cytoplasmic distribution of NBCe1‐A‐EGFP. C, cells transfected with Q913R‐EGFP also exhibit cytoplasmic distribution of NBCe1‐A‐EGFP. Images in (A) to (C) are representative of the distribution of NBCe1‐A‐EGFP as observed in more than four cells in each of three independent transfections. The brightness of all images in (A) to (C) has been increased by 50% for clarity. The thin white lines in each indicate the location of the optical slices displayed in the other planes shown in each panel. D, Pearson's coefficients report the degree of co‐localization of EGFP and Na,K‐ATPase immunoreactivity in five images equivalent to those shown in (A) to (C). *Significance vs. WT‐EGFP with 95% confidence using an ANOVA with Tukey's comparison test. n.s., no significant difference.