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Figure 3


Fig. 3. Analysis of the PI4KIIIß-14-3-3 interaction by BiFC with fluorescence flow cytometry and confocal microscopy. (A) HEK293 cells were cotransfected with plasmids encoding wildtype YN-PI4KIIIß/YC-14-3-3{tau} (red) or YN-PI4KIIIß/YC-DN-14-3-3{tau} (blue). Cells were harvested 48 hours after transfection in PBS supplemented with 5% FCS and 0.05% sodium azide and BiFC was analysed by fluorescence flow cytometry. Untransfected cells were used as a negative control (filled). The data are presented in a histogram depicting EYFP fluorescence (FL1) (x axis) vs cell number (count) (y axis). The results are representative of three independent experiments. (B) COS7 cells were cotransfected with YN-PI4KIIIß and YC-14-3-3{tau}. Cells were fixed two days after transfection, stained with anti-p230 followed by anti-mouse IgG Cy5. BiFC is shown in green, p230 proteins in red. Shown is a single optical section. Bar, 10 µm.