Figure 4.

FRET measurements of TC45 and TC45^E/Ain cells display both intramolecular and intermolecular interaction.A, left, representative FACS plots for control, CFP-linker-YFP, CFP-TC45-YFP, and CFP-TC45^E/A-YFP. Top, double positive cells for CFP and YFP expression. Middle, selection of FRET-positive cells excluding the false-positive signal because of direct YFP excitation. Bottom panel, triangular gating containing FRET-positive cells. Right, bar graphs summarizing the results from the panels on the left. Representative examples of the FACS plots used to determine the gating strategy for these measurements are shown in Fig. S6A. B, left, three panels are representative FACS plots for the overexpression of NLS-CFP and ^NLS-YFP, CFP-TC45-CFP and YFP-TC45-YFP, CFP-TC45^E/A-CFP and YFP-TC45^E/A-YFP. Top, middle, and bottom panels are as described above. Right, bar graphs summarizing the results from the panels on the left. Representative examples of the FACS plots used to determine the gating strategy for these measurements are shown in Fig. S6B. Data represent seven replicates and were analyzed with Kruskal–Wallis test (∗p < 0.05; ∗∗p < 0.01). CFP, cyan fluorescent protein; FACS, fluorescence-activated cell sorting; NLS, nuclear localization signal.