TABLE 2.
FCCS fitting parameters (± S.E.) for the GFP- and mCherry-tagged GPCRs in the plasma membrane of transiently transfected HEK 293 cells
Decay times (τD, 1 (ms); τD, 2 (ms); fractions (τD, 1 (%); τD, 2 (%)), and CC values (G(0)x/G(0)min) are listed. The best fits were obtained assuming free diffusion in two dimensions and two components (2D_2C). Triplet fractions (TF (%)) ranged from 20 to 50% and triplet decay times (τD,F (μs)) from 20 to 200 μs.
| GPCRs | τD, 1 | τD, 2 | τD, 1 | τD, 2 | N | G(0)x/G(0)min |
|---|---|---|---|---|---|---|
| % | % | ms | ms | % | ||
| CRF1 | ||||||
| GFP | 40 ± 1 | 60 ± 1 | 0.43 ± 0.02 | 36 ± 1 | 108 | 17.2 ± 0.5 |
| mCherry | 56 ± 1 | 44 ± 1 | 0.15 ± 0.01 | 39 ± 2 | ||
| CRF2(a)R | ||||||
| GFP | 51 ± 1 | 49 ± 1 | 0.57 ± 0.03 | 37 ± 1 | 93 | 9.3 ± 0.3 |
| mCherry | 63 ± 1 | 37 ± 1 | 0.16 ± 0.01 | 38 ± 2 | ||
| TSHR | ||||||
| GFP | 33 ± 1 | 67 ± 1 | 0.33 ± 0.03 | 34 ± 1 | 98 | 11.4 ± 0.4 |
| mCherry | 58 ± 1 | 42 ± 1 | 0.11 ± 0.01 | 33 ± 1 | ||
| LHR | ||||||
| GFP | 53 ± 3 | 47 ± 3 | 0.29 ± 0.02 | 25 ± 2 | 42 | 16.3 ± 1.1 |
| mCherry | 60 ± 2 | 40 ± 2 | 0.13 ± 0.03 | 22 ± 3 | ||
| V2R | ||||||
| GFP | 41 ± 1 | 59 ± 1 | 0.46 ± 0.03 | 37 ± 2 | 117 | 14.6 ± 0.5 |
| mCherry | 60 ± 1 | 40 ± 1 | 0.13 ± 0.01 | 38 ± 2 | ||
| ETBR | ||||||
| GFP | 46 ± 2 | 54 ± 2 | 0,26 ± 0,02 | 23 ± 1 | 86 | 14.0 ± 0.6 |
| mCherry | 66 ± 1 | 34 ± 1 | 0.08 ± 0.01 | 25 ± 2 | ||
| PAR1 | ||||||
| GFP | 64 ± 2 | 36 ± 2 | 0.27 ± 0.01 | 28 ± 1 | 84 | 14.0 ± 0.5 |
| mCherry | 73 ± 1 | 27 ± 1 | 0.15 ± 0.01 | 23 ± 1 | ||