Immunofluorescence imaging of the p.G40S and p.R244P mutant forms of Cx47. As shown, cells transfected with p.G40S alone or with separate constructs for p.G40S and for p.R244P show GJ plaques (white arrows). Note that the cells expressing both p.G40S and p.R244P showed a trend toward less plaques than WT or p.G40S alone, consistent with the reduced amount of p.G40S DNA used to transfect the p.G40S+p.R244P-expressing cells. On the other hand, no plaques were seen with the p.R244P mutant when expressed alone. The control micrograph is of N2a cells transfected with pIRES2-EGFP and then immunostained with anti-Cx47, so the green should represent the background fluorescence of our ICC protocol for labeling Cx47. In the bottom right panel, significance was determined using pairwise Fisher’s exact tests with Šídák modification.