Extended Data Fig. 10: SmFRET-RAP data for Sf-SecR and Sf-MOR.
From: Single-molecule FRET imaging of GPCR dimers in living cells
a, Acceptor (top) and donor (bottom) labeled receptor densities before photobleaching for smFRET-RAP (left panel) compared to those used for smFRET at lower receptor expression levels (right panel) reproduced from Extended Data Fig. 6b. Dots represent the number of acceptor (nAcc) or donor (nDon) particles per area for single cells. Box plot details are described in the legend of Extended Data Fig. 2b. b, c, TIRF images of representative CHO cells expressing labeled b, Sf-SecR from 7 cells and (c) Sf-MOR from 7 cells before donor and acceptor photobleaching (left panel), ~30 seconds after photobleaching (middle panel), and ~2 – 3 minutes after photobleaching (right panel) showing the recovery of labeled receptors (scale bar, 5 μm). d, Representative trajectories and sensitized acceptor intensity time traces for Sf-SecR. The top trajectory and trace are derived from the image sequence shown in Fig. 5b. e, Duration of smFRET events of SecR interactions determined from the tracking duration of sensitized acceptor trajectories. The distribution comprised of 4,232 trajectories from 21 cells was fit to a single exponential with decay constant (τ).
