Fig. 7: ^nirCATCHFIRE: a fluorogenic chemically induced dimerization tool with NIR fluorescence readout.

a Schematic illustrating the fluorogen-induced interaction between EGFP-^FIREtag and Tom20-ECFP-^nir-FIREmate for chemically induced recruitment at the outer mitochondrial membrane. b–e HeLa cells co-expressing EGFP-^FIREtag together with Tom20-ECFP-^nir-FIREmate were treated with 10 µM of either HPAR-3OM (b, c) or HPAR−3,5DOM (d, e), and imaged by timelapse confocal microscopy. b, d Representative confocal micrographs before and after the addition of the fluorogen (see also Supplementary Movies 5, 6). Scale bar 10 µm. c, e Temporal evolution of the mitochondrial EGFP and ^nirCATCHFIRE fluorescence of (c) n = 15 cells (from three independent experiments) and (d) n = 14 cells (from four independent experiments). The dashed line indicates fluorogen addition. f Schematic illustrating how fluorogen-induced interaction between LAMP1–mCherry–^FIREtag and ^nir-FIREmate–KIF17 allows the chemically induced anterograde transport of lysosomes. g HeLa cells co-expressing LAMP1–mCherry–^FIREtag and ^nir-FIREmate–KIF17 were imaged by spinning disk microscopy for 30 min, then HPAR−3,5DOM was added. Representative micrographs of each step (see also Supplementary Movie 7). Experiments were repeated three times with similar results. Scale bars 10 µm. h Kymograph showing the LAMP1–mCherry–^FIREtag along the yellow line in (e) over time. See Supplementary Table 8 for detailed imaging settings. Source data are provided as a Source Data file.