Table 1.
Results of FLIM-FRET analysis and co-localization measurement (Pearson's correlation coefficient).
| Donor protein | Acceptor protein | Pearson's correlation coefficient ± SD | Donor lifetime [τD (ns) ± SD] | Donor lifetime in the presence of potential acceptor [τ DA (ns) ± SD] | FRET efficiency (%) |
|---|---|---|---|---|---|
| GFP | RFP | 0.78 ± 0.03 | 2.51 ± 0.01 | 2.48 ± 0.01 | 1.19 |
| PCNA2_GFP | RFP_PCNA2 | 0.77 ± 0.01 | 2.51 ± 0.01 | 2.39 ± 0.01 | 4.78 |
| CycD1;1_GFP | RFP_PCNA1 | 0.83 ± 0.01 | 2.38 ± 0.02 | 2.37 ± 0.02 | 0.42 |
| CycD1;1_GFP | RFP_PCNA2 | 0.84 ± 0.01 | 2.38 ± 0.02 | 2.36 ± 0.01 | 0.84 |
| CycD2;1_GFP | RFP_PCNA1 | 0.71 ± 0.09 | 2.45 ± 0.01 | 2.41 ± 0.01 | 1.63 |
| CycD2;1_GFP | RFP_PCNA2 | 0.70 ± 0.04 | 2.45 ± 0.01 | 2.43 ± 0.02 | 0.82 |
| CycD3;1_GFP | RFP_PCNA1 | 0.92 ± 0.02 | 2.45 ± 0.02 | 2.44 ± 0.02 | 0.41 |
| CycD3;1_GFP | RFP_PCNA2 | 0.92 ± 0.01 | 2.45 ± 0.02 | 2.44 ± 0.03 | 0.41 |
| CycD3;2_GFP | RFP_PCNA1 | 0.86 ± 0.01 | 2.49 ± 0.01 | 2.48 ± 0.00 | 0.40 |
| CycD3;2_GFP | RFP_PCNA2 | 0.84 ± 0.05 | 2.49 ± 0.01 | 2.48 ± 0.01 | 0.40 |
| CycD3;3_GFP | RFP_PCNA1 | 0.80 ± 0.02 | 2.43 ± 0.04 | 2.42 ± 0.02 | 0.41 |
| CycD3;3_GFP | RFP_PCNA2 | 0.90 ± 0.02 | 2.43 ± 0.04 | 2.40 ± 0.02 | 1.23 |
| CycD4;1_GFP | RFP_PCNA1 | 0.80 ± 0.01 | 2.53 ± 0.01 | 2.43 ± 0.02 | 3.95 |
| CycD4;1_GFP | RFP_PCNA2 | 0.71 ± 0.02 | 2.53 ± 0.01 | 2.48 ± 0.02 | 1.98 |
| CycD4;2_GFP | RFP_PCNA1 | 0.70 ± 0.00 | 2.53 ± 0.01 | 2.44 ± 0.02 | 3.56 |
| CycD4;2_GFP | RFP_PCNA2 | 0.74 ± 0.01 | 2.53 ± 0.01 | 2.40 ± 0.01 | 5.14 |
| CycD5;1_GFP | RFP_PCNA1 | 0.83 ± 0.01 | 2.32 ± 0.02 | 2.31 ± 0.01 | 0.43 |
| CycD5;1_GFP | RFP_PCNA2 | 0.85 ± 0.02 | 2.32 ± 0.02 | 2.30 ± 0.03 | 0.86 |
| CycD6;1_GFP | RFP_PCNA1 | 0.85 ± 0.03 | 2.44 ± 0.01 | 2.34 ± 0.02 | 4.10 |
| CycD6;1_GFP | RFP_PCNA2 | 0.82 ± 0.04 | 2.44 ± 0.01 | 2.37 ± 0.02 | 2.87 |
| CycD7;1_GFP | RFP_PCNA1 | 0.93 ± 0.00 | 2.40 ± 0.01 | 2.25 ± 0.02 | 6.25 |
| CycD7;1_GFP | RFP_PCNA2 | 0.86 ± 0.01 | 2.40 ± 0.01 | 2.31 ± 0.02 | 3.75 |
| CycD1;1_GFP | PCNA1_RFP | 0.87 ± 0.02 | 2.48 ± 0.01 | 2.45 ± 0.02 | 1.13 |
| CycD1;1_GFP | PCNA2_RFP | 0.87 ± 0.01 | 2.48 ± 0.01 | 2.49 ± 0.02 | 0.08 |
| CycD2;1_GFP | PCNA1_RFP | 0.79 ± 0.05 | 2.57 ± 0.01 | 2.54 ± 0.02 | 1.17 |
| CycD2;1_GFP | PCNA2_RFP | 0.77 ± 0.04 | 2.57 ± 0.01 | 2.54 ± 0.01 | 0.94 |
| CycD3;1_GFP | PCNA1_RFP | 0.90 ± 0.00 | 2.52 ± 0.01 | 2.48 ± 0.01 | 1.75 |
| CycD3;1_GFP | PCNA2_RFP | 0.89 ± 0.01 | 2.52 ± 0.01 | 2.50 ± 0.01 | 0.87 |
| CycD3;2_GFP | PCNA1_RFP | 0.82 ± 0.03 | 2.51 ± 0.01 | 2.49 ± 0.01 | 0.72 |
| CycD3;2_GFP | PCNA2_RFP | 0.83 ± 0.02 | 2.51 ± 0.01 | 2.48 ± 0.01 | 1.04 |
| CycD3;3_GFP | PCNA1_RFP | 0.84 ± 0.01 | 2.53 ± 0.01 | 2.49 ± 0.01 | 1.50 |
| CycD3;3_GFP | PCNA2_RFP | 0.86 ± 0.03 | 2.53 ± 0.01 | 2.49 ± 0.01 | 1.42 |
| CycD4;1_GFP | PCNA1_RFP | 0.93 ± 0.01 | 2.54 ± 0.03 | 2.49 ± 0.01 | 2.13 |
| CycD4;1_GFP | PCNA2_RFP | 0.91 ± 0.01 | 2.54 ± 0.03 | 2.50 ± 0.02 | 1.50 |
| CycD4;2_GFP | PCNA1_RFP | 0.86 ± 0.02 | 2.52 ± 0.02 | 2.50 ± 0.01 | 0.48 |
| CycD4;2_GFP | PCNA2_RFP | 0.80 ± 0.03 | 2.52 ± 0.02 | 2.50 ± 0.02 | 0.48 |
| CycD5;1_GFP | PCNA1_RFP | 0.85 ± 0.01 | 2.40 ± 0.03 | 2.39 ± 0.03 | 0.50 |
| CycD5;1_GFP | PCNA2_RFP | 0.86 ± 0.01 | 2.40 ± 0.03 | 2.40 ± 0.04 | 0.08 |
| CycD6;1_GFP | PCNA1_RFP | 0.87 ± 0.01 | 2.49 ± 0.01 | 2.46 ± 0.02 | 0.57 |
| CycD6;1_GFP | PCNA2_RFP | 0.87 ± 0.02 | 2.49 ± 0.01 | 2.48 ± 0.01 | 0.08 |
| CycD7;1_GFP | PCNA1_RFP | 0.92 ± 0.01 | 2.49 ± 0.02 | 2.43 ± 0.01 | 1.94 |
| CycD7;1_GFP | PCNA2_RFP | 0.90 ± 0.01 | 2.49 ± 0.02 | 2.44 ± 0.02 | 1.53 |
(τ D) and (τ DA) represent the average fluorescence lifetimes of donor proteins and donor proteins in the presence of a potential acceptor respectively. SD, standard deviation. The results are an average of three independent experiments.