Figure 4—figure supplement 1.

Figure 4—figure supplement 1.

(A) Confocal image of through an immunocytochemical staining of a HEK293T cell transfected with zebrafish kcnk5b-GFP transgene (B) DAPI staining of the same cell in panel A. (C) Merged panels A and B. (D–L) Serial confocal cross sections for GFP in a live HEK293T cell from its surface (D) through to a mid-section of the cell (L) after transfection with the zebrafish kcnk5b-GFP transgene. (M) FRET-FLIM image after measuring the life time of CFP of the K⁺ FRET reporter KIRIN in a cell transfected with a control plasmid only expressing mCherry. The inset image of the confirmation of the FRET reporter depicting how FRET occurs. (N) FRET-FLIM image after measuring the life time of CFP of the K⁺ FRET reporter KIRIN in a cell transfected with a plasmid expressing kcnk5b-mCherry. The inset image of the confirmation of the FRET reporter depicting how FRET do not occur. (O) The graphed assessments of the changes in CFP lifetime in the control mCherry-expressing cells and the cells expressing kcnk5b-mCherry. The diagrams to the right of the graph depict the conformational changes that result in the values of the lifetimes of CFP. The data for each experiment represents three or more experiments. The graphed data points show all technical replicates.