Fig. 5.

Cytosolic mEo3.2 displays reversible photobleaching in HEK 293T cells. A: representative fluorescence recovery after photobleaching (FRAP) images (prebleach, bleach, and recovery) of HEK 293T cells expressing mEos3.2. Scale bar = 10 μm. B: quantification of FRAP images. FRAP images were acquired every 30 min for 9 h. Pixel intensities of the region of interest (ROI), cell, and background were acquired. Background was subtracted from ROI and whole cell. Images were normalized to prebleach intensities. Data were fitted into a one-phase association equation to extrapolate a reversible photoswitching rate (n = 3 cells). C: mobile fractions of fixed mEos3.2-expressing HEK 293T cells. AU, arbitrary units.