Fig. 6: Visualization of ternary protein interactions at the junctions of organelles.

a–c HeLa cells co-expressing pFAST_99-114–^NFRB–ECFP–CAAX, Tom20-mCherry–^CFRB–pFAST_1-98 and FKBP–pFAST_115-125 were treated with 10 μM HBR-2,5DM. Cells were imaged by time-lapse confocal microscopy after addition of 500 nM of rapamycin. Experiments were repeated three times with similar results. b Representative micrographs before and after addition of rapamycin (see also Supplementary Movie 5). c Correlation of the ECFP and mCherry signals before and after rapamycin addition. d–f HeLa cells co-expressing pFAST_99–114–^NFRB–ECFP–Cb5, Lyn11-mCherry–^CFRB–pFAST_1–98 and FKBP–pFAST_115–125 were treated with 10 μM HBR-2,5DM. Cells were imaged by time-lapse confocal microscopy after addition of 500 nM of rapamycin. Experiments were repeated three times with similar results. e Representative micrographs before and after addition of rapamycin (see also Supplementary Movie 6). f Correlation of the ECFP and mCherry signals before and after rapamycin addition. g–i HeLa cells co-expressing pFAST_99–114–^NFRB–ECFP–Cb5, Tom20-mCherry–^CFRB–pFAST_1–98 and FKBP–pFAST_115–125 were treated with 10 μM HBR-2,5DM. Cells were imaged by time-lapse confocal microscopy after addition of 500 nM of rapamycin. Experiments were repeated three times with similar results. h Representative micrographs before and after addition of rapamycin (see also Supplementary Movie 7). i Correlation of the EGFP and mCherry signals before and after rapamycin addition. j–l HeLa cells co-expressing pFAST_99-114–^NFRB–ECFP–Cb5, Lyn11-mCherry–^CFRB–pFAST_1-98, and Tom20–FKBP–pFAST_115-125 were treated with 10 μM HBR-2,5DM. Cells were imaged by time-lapse confocal microscopy after addition of 500 nM of rapamycin. Experiments were repeated three times with similar results. k Representative micrographs before and after addition of rapamycin (see also Supplementary Movie 8). l Correlation of the EGFP and mCherry signals before and after rapamycin addition. c, f, i, l Data represent the mean of n = 10 (c), 9 (f), 10 (i), 11 (l) from three independent experiments. Two-tailed Student’s t-test assuming equal variance was used to compare correlations before and after addition of rapamycin: (c) t(8) = 5.14, P value < 0.0001; (f) t(9) = 9.527, P value = 0.0009; (i) t(9) = 9.991, P value < 0.0001, (l) (f) t(10) = 5.808, P value = 0.0002. Scale bars 20 μm. Source data are provided as a Source Data file.