Supplementary Figure 6: FLIRT-targeted control embryos maintain cell polarity and viability.

a, Schematic (left) and representative images (right) depicting an assay to monitor the effect of equatorial FLIRT on cell polarity and daughter size asymmetry in one-cell embryos expressing PAR-6 (PARtitoning defective) and PAR-2. Quantification of cell polarity and daughter cell asymmetry during cell division (bottom center) and at the two-cell stage (bottom right). N = 7 biologically independent embryos for 16 °C (0 mW), 7 for 16 °C (8.5 mW), 6 for 14 °C (0 mW), and 7 for 14 °C (10.1 mW). Unpaired two-tailed t-test; n.s., no significance, P > 0.05. Error bars, mean +/− s.d.; see Supplementary Table 1 for statistical analysis. b, Experimental timeline for post-FLIRT viability assay. The yellow arrowhead indicates embryo transfer to a worm plate, gray arrowheads indicate developmental imaging, and yellow arrows indicate embryo location on a worm plate. Images of a representative individual control embryo rescued after equatorial FLIRT (top row—and shown at the two-cell stage in the micrograph indicating the FLIRT ROI) or dissected at the one-cell stage (no FLIRT control, bottom) throughout development. Quantification of embryonic viability for isolated one-cell-stage embryos (right). Red (schematic) and white (images) dashed circles indicate FLIRT-targeted ROIs. Time is in minutes after FLIRT initiation. The number of biologically independent experimental replicates (N) is indicated on individual bar graphs. White scale bars, 10 µm; yellow scale bar, 100 µm.