Figure 1:

FLIRT calibration and application for spatiotemporal control of ts protein function in vivo

(a) Fast-acting ts mutant proteins are rapidly inactivated upon temperature upshift^{3, 6}. (b) Schematic of FLIRT targeting in which an IR laser is used to locally inactivate ts mutant protein function. (c) Experimental schematic (left) and representative time lapse images (right) of cell-specific FLIRT targeting in 2-cell C. elegans embryos. See Supplementary Video 1. (d) Experimental schematic (left) and representative time lapse images (right) of subcellular FLIRT targeting either an equatorial or polar region in myosin-II(ts) 1-cell embryos. Embryos were FLIRT-targeted either throughout division (top 3 rows, see Supplementary Video 3) or for ~8 min to test reversibility (bottom row, see Supplementary Video 5). The number of AB and P1 cells (c) or 1-cell embryos (d) from biologically independent embryos that successfully completed cell division is indicated below each experimental schematic (left). Red (schematic) or white (images) dashed circles indicate the FLIRT-targeted regions. Time is in sec after FLIRT initiation. Scale bars=10 μm.