M93I-GFP traffics normally to the plasma membrane and co-localizes with Cx26WT-mCherry. HeLa Ohio cells were transfected with Cx26WT-mCherry (a, red), M93I-GFP (b, green), or both Cx26WT-mCherry and M93I-GFP (c), and protein localization was analyzed after 24 hours. In both single and double transfected cells, Cx26WT-mCherry and M93I-GFP expression was observed both within the cytoplasm (white arrow heads) and at the plasma membrane (white arrows), indicating cellular stores and connexin (Cx) trafficking to the membrane. Cx26WT-mCherry and M93I-GFP co-localized throughout the cell (yellow) and formed visible gap junction plaques at points of cell-to-cell contact. Nuclei are stained with 4′-6-diamidino-2-phenylindole (DAPI; blue). Bar=20 μm.