Fig. 4: Evaluation of tripartite split systems.
a–f Normalized average fluorescence of about 20,000 HEK293T cells co-expressing the pairs RspAFAST99-114–FRB–RspAFAST1-98 / FKBP–RspAFAST115-125 (a–c) or pFAST99-114–FRB–pFAST1-98 / FKBP–pFAST115-125 (d–f) treated without or with 500 nM of rapamycin, and with 0, 1, 5, 10, 25 or 50 μM of HMBR (b, e) or HBR–2,5DM (c,f). Data represent the mean ± SD of three independent experiments (b, f) and two independent experiments (c, e). g–i HEK293T cells expressing pFAST99-114–FRB–pFAST1-98 and FKBP–pFAST115-125 were treated with 5 μM HBR-2,5DM. Cells were imaged after addition of 500 nM of rapamycin. Experiments were repeated three times with similar results. g Representative micrographs before and after addition of rapamycin (see also Supplementary Movie 2). Scale bars 20 μm. h Temporal evolution of the fluorescence signal (F/Fmax) after addition of rapamycin n = 26 cells from three independent experiments. F is the fluorescence intensity at time t, and Fmax is the maximal fluorescence intensity reached during the time-lapse. i Fluorescence fold increase (Fmax/Fmin) upon addition of rapamycin. Fmin is the fluorescence intensity at time t = 0, and Fmax is the maximal fluorescence intensity reached during the time-lapse. h, i Each cell is color-coded according to the biological replicate it came from. h The dot lines represent the mean value of each biological replicate, while the black line represents the mean of the three biological replicates. i The solid circles correspond to the mean of each biological replicate. The black line represents the mean ± SD of the three biological replicates. Source data are provided as a Source Data file.
