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. 2012 Dec 6;288(6):4378–4388. doi: 10.1074/jbc.M112.388629

FIGURE 3.

FIGURE 3.

Calculation of apparent FRET efficiency for CFP/YFP-tagged constructs. A, emission spectra of untransfected cells (dashed lines), 6.2C:6.2Y (1:1) co-transfected cells (thin solid lines), excited at 430 (blue) or 480 nm (yellow), and specific FP-associated fluorescence obtained by subtraction of non-FP fluorescence (thick solid lines). B, specific FP fluorescence of 6.2C (blue) or 6.2Y (yellow) excited at 430 nm (dashed) or 480 nm (solid). 525-nm 6.2Y fluorescence excited by 430 nm (Eapp) was ∼6% of that excited by 480 nm. C, specific FP fluorescence of co-expressed 6.2C:6.2Y (1:1) excited at 430 nm (blue) or 480 nm (yellow), scaled 6.2C fluorescence excited at 430 nm (see text), and subtracted 6.2Y-specific emission. 525 nm 6.2Y fluorescence excited by 430 nm (Eapp) was ∼22% of that excited by 480 nm.