DP disease mutations impair Cx43 membrane localization. (A) Immunostaining of Pg, GFP, and Cx43 in NHEKs expressing WT and mutated DPII-GFP constructs. Pg staining was used to mark regions of cell–cell contact between cells expressing DPII-GFP constructs (white arrows). Regions were overlaid on thresholded Cx43 images for quantification. Bars, 10 µm. (B) Quantification of Cx43 fluorescence intensity at cell–cell contacts in NHEKs (i) and HL-1s (ii; n = 9 image fields from three independent experiments). (C) Quantification of cytoplasmic GFP intensity in NHEKs expressing WT and mutated DPII-GFP constructs (n = 9 image fields from three independent experiments). Error bars indicate SEM.