Extended Data Fig. 8: Kinetics of association and dissociation of split-greenFAST and split-redFAST.
From: Orthogonal fluorescent chemogenetic reporters for multicolor imaging
a, b, Representative micrographs (n = 3 from 3 experiments) of HEK293T cells co-expressing FK506-binding protein (FKBP) fused to CFAST11 and FKBP-rapamycin-binding domain of mammalian target of rapamycin (FRB) fused to either greenNFAST (a) or redNFAST (b) were labeled with both 5 μM HMBR and 10 μM HBR-3,5DOM, and imaged before and after addition of 500 nM rapamycin. The green channel shows HMBR fluorescence, while the magenta channel shows HBR-3,5DOM fluorescence. Graphs show the temporal evolution of the fluorescence intensity of individual cells (n = 7 and 8) after rapamycin addition. c, d, Representative micrographs (n = 3 from 3 experiments) of HEK293T cells co-expressing FKBP fused to CFAST11 and FBBP fused to either greenNFAST (c) or redNFAST (d) treated with 100 nM AP1510 and labeled with both 5 μM HMBR and 10 μM HBR-3,5DOM. Cells were then imaged before and after the addition of 1.1 μM rapamycin. The green channel shows HMBR fluorescence, while the magenta channel shows HBR-3,5DOM fluorescence. Graphs show the temporal evolution of the fluorescence intensity of individual cells (n = 8 and 11) after rapamycin addition.
