Fig. 7
Live-cell recordings of cellular organelles labeled with GCE-tag and a tetrazine-conjugated Fl-dye. a Live-cell recordings of cells expressing GCE-tag-GFP-SKL labeled with SiR-Tet. Upper panel: maximum projection images of the entire cell taken at time 0. Bottom panel: sequential zoomed-in images of a subset of the cell (corresponds to square in upper panel) taken from the movie series. Every fourth frame from the movie series is shown (Additional file 4: Movie S3). b Live-cell recordings of cells expressing GCE-tag-CD63 labeled with TAMRA-Tet. Left, a maximum projection image of the entire cell taken at time 0. Sequential zoomed-in images of a subset of the cell (corresponds to square in left panel) taken from the movie series are shown to the right. Every fourth frame from the movie series is shown (Additional file 6: Movie S5). c FRAP analysis of cells expressing GCE-tag-ERcb5TM labeled with TAMRA-Tet. Snapshots of a subset of a representative cell taken from the movie series (Additional file 8: Movie S7) are shown in the upper panel. Sequential images of the bleached ROI are shown in the bottom panel (Additional file 9: Movie S8). Plot on the right represents the exponential fit of fluorescence intensity recovery versus time after photobleaching in the ROI. Intensity values were corrected for unintentional bleaching and normalized to intensity levels measured pre-bleaching. Results presented in each panel were obtained in at least 3 independent experiments. Scale-bars, 10 μm; zoomed-in images, 2 μm