Fig. S5

Knockout of miR-126a impairs trunk lymphatic network formation. (A) Assay of the deletion efficiency of Cas9 with two gRNAs pairs. (B) Confocal images of trunk vessels in 55 hpf embryos that were injected either with Cas9+gRNAs, or Cas9 alone. (C) Quantification of PLs in control and mosaic miR-126a^KO embryos at 55 hpf. (D) Confocal images of TD in 120 hpf embryos that were either injected with Cas9+gRNAs, or Cas9 alone. (E) Identify of miR-126a^KO F2 embryos with PCR and sequencing. Sequence with deletions induced by Cas9/gRNAs is indicated in right panel. WT: wild type; mut: mutant. (F) Confocal images of PL formation from 40 to 52 hpf. The red arrows indicate normal PLs development and the whites denote defective PL formation. (G) Bright-field side views of the gross morphology in 120 hpf embryos. Red arrowhead indicates edema. (H) Confocal images of TD in 120 hpf embryos. The red arrows denote normal TD. (I) A graph represents the genotype distribution of F2 embryos with PL defect. Scale bars, 100 μm.