Supplementary Figure 2: Analysis of the gene expression patterns of itgb1b, tspan4a and tspan7 in zebrafish embryos by whole-mount in situ hybridization.
From: Migrasomes provide regional cues for organ morphogenesis during zebrafish gastrulation
a. Expression pattern of itgb1b viewed laterally and from the animal pole. Scale bar, 100 μm. b. Representative images of migrasomes in WT embryos and itgb1b mutants. c. Images of gastrulas from three independent experiments were pooled and quantified for the number of migrasomes. WT, n=19 embryos; itgb1b-/-, n=26 embryos, ***P<0.001. Data represent mean ± s.e.m, P values were calculated using a two-tailed, unpaired t-test. d. Quantification of the average speed of cell migration in WT embryos and itgb1b mutants. 4D images of gastrulas were acquired by light-sheet microscopy and the average migration speed was measured by Imaris software. n=3 embryos in WT, n=4 embryos in itgb1b-/-. ns, P>0.05. Data represent mean ± s.e.m, P values were calculated using a two-tailed, unpaired t-test. Data were pooled from two independent experiments. e-f. Expression pattern of tspan4a (e) and tspan7 (f) viewed laterally and from the animal pole. Scale bar, 100 μm. g. The efficiency of morpholinos was tested using reporters carrying the gene-specific 5’UTR upstream of GFP. For each treatment, seven embryos were imaged simultaneously with GFP fluorescence and bright field. Scale bar, 1 mm. h, i. Quantification of the number of migrasomes after morpholino injection. Control and gene-specific MOs were injected at the one-cell stage (10 ng per embryo). Migrasomes were counted at the shield stage. (h) Ctrl MO, n=10; tspan4a MO, n=10 embryos; *P<0.05. (i) Ctrl MO, n=11 embryos; tspan7 MO, n=13 embryos; ***P<0.001. Data represent mean ± s.e.m, P values were calculated using a two-tailed, unpaired t-test. j. Whole-mount in situ hybridization of 1-cell-stage wild-type, MZtspan4a and MZtspan7 embryos. 19 embryos were imaged simultaneously in each group. Scale bar, 1 mm. The experiments in a, e, f were done twice. The experiments in g-j were done once. The numerical source data for c,d,h and i are shown in Supplementary Table 3.
