Extended Data Fig. 1: miR-128^Δ/Δ phenotype in vascular and hematopoietic development.

(a) Genomic location of miR-128-1 and miR-128-2 and qRT-PCR analysis of r3hdm1 and arpp21 (3 independent pools of ~10 embryos; Two-tailed Mann-Whitney test). (b) Representative images showing cmyb and lcp1 cells. (c) Time-lapse with dragon-tail analysis of WT and 128^Δ/Δ Tg(kdrl:mCherry^s896,cmyb:GFP^zf169) from ~24 to 50 hpf. The color scale represents the time a cell to delaminate. (d) Confocal imaging of WT and 128^Δ/Δ embryos at 27 hpf. (e) Percentage of ECs over total number of cells in WT (n = 11 embryos) and 128^Δ/Δ (n = 10 embryos) at 26 hpf by flow cytometry (3 individual experiments, Two-tailed Mann-Whitney test). (f) WISH of WT and 128^Δ/Δ with notch3 and flt4. (g) Images of WT and 128^Δ/Δ embryos. (h) WISH of etv2 and scl at 12 hpf. (i) qRT-PCR analysis of etv2 and scl (8 to 9 independent pools of ~10 embryos) at 12 hpf (Two-tailed Mann-Whitney test). (j) WISH of rag1+ cells at 4.5 dpf (n = WT, 36, 128^Δ/Δ, 39 embryos; p-value = 0.001; 3 independent experiments, Mann-Whitney test). (k) O-dianisidine staining at 6 dpf (n = WT, 38, 128^Δ/Δ, 44 embryos; p-value < 0.0001; 3 independent experiments, Mann-Whitney test). (l) Sudan black staining at 6 dpf (n = WT, 32, 128^Δ/Δ, 27 embryos; 3 independent experiments, Two-tailed Mann-Whitney test). (m) O-dianisidine staining of WT and 128^Δ/Δ at ~2.5 dpf. (n) Sudan black staining of WT and 128^Δ/Δ at ~2 dpf. (o) qRT-PCR analysis of miR-128 normalized with U6 (n = MO-ctrl, 3, MO-128; p-value = 0.0224; 3 independent pools of ~10 embryos, Two-tailed one sample t and Wilcoxon test). (p) Flow cytometry population analysis of ~1-month-old dissected whole KM zebrafish injected with MO-control or MO-128. All quantifications are represented with mean ± SEM. ns: p > 0.05, *p ≤ 0.05. Abbreviations: aorta gonad mesonephros (AGM), dorsal aorta (DA), posterior cardinal vein (PCV), caudal hematopoietic tissue (CHT), thymus (T), intersegmental vessel (ISV) and (kidney marrow (KM).