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Fig. 1. A u-adFRET assay. Keratinocytes were co-transfected with (wt)Cx30-ECFP (cyan pseudocolour, donor) and (wt)Cx26-EYFP (yellow pseudocolour, acceptor). (A) Expression patterns of (wt)Cx30-ECFP (left), (wt)Cx26-EYFP (middle) and the merged image (right) before bleaching (top row) and the changes of fluorescence intensity in areas of interest (c to h) after bleaching (bottom row) show enhanced fluorescence intensity of donor (cyan pseudocolour, left) and reduced fluorescence intensity of acceptor (yellow pseudocolour, middle). Cyan and yellow pseudocolour look-up tables were designed to produce white in the merged image when perfectly colocalised. Areas labelled a and b were used as a background reference and an unbleached reference respectively. (B) Example profiles (from region g) of relative expression concentrations of donor and acceptor (top) and the changes of fluorescence intensity in donor and acceptor before and after bleaching (bottom). Bar, 10 µm.





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