Fig. 2: Validating binary-FRET.

a Cartoon depicting fluorescent protein control constructs. b In vitro fluorescence lifetime decays, c In vitro time-resolved, fluorescence anisotropy decays. Steady-state anisotropy and fluorescence lifetime images (128 × 128 pixels corresponding to 42.6 µ × 42.6 µ) of HEK cells expressing ChVV (d), UrVV (e), ChVA (f) and UrVA (g). Anisotropy color bar ranged from 0.3 (dark red) to 0.5 (dark purple). Lifetime color bars are in picoseconds and ranged from 0 ps (dark red) to 3500 ps (dark purple). Source data are provided as a Source Data file.