Splice-blocking dachb-morpholino (dachb-MO-S) knockdown inhibits the development of the pancreatic islet- and β-cells. Embryos were (a) uninjected (control) or injected at the one-cell stage with (b) 4 ng standard MO (Std. MO; control), (c) 4 ng p53-MO (control) or (d,e,f) a combination of p53-MO and dachb-MO-S at (d) 1 ng + 1ng, (e) 2 ng + 2 ng or (f) 4 ng + 4 ng. (ai-fi) Bright-field images were acquired at 30 h post-fertilization (hpf), after which the embryos were whole-mount double-immunolabeled with antibodies to insulin (green) and islet 1 and 2 (red) to show the localization of the β-cells and islet cells in the endocrine pancreas, respectively. (aii–fii) These are all single optical sections taken through the middle of pancreatic islet. (g–h) The number of (g) β-cells and (h) islet cells, and (i,j) the volume of (i) β-cells and (j) islet cells. (k) The circularity of the pancreatic islet. (l) Bar charts showing the numbers of (li) β-cells and (lii) islet cells in embryos at 24 hpf and 30 hpf after they were injected at the one-cell stage with either p53-MO (controls) or p53-MO and dachb-MO-S (at the amounts described above). The data are presented as the mean ± standard error of the mean of n = 7 to 41 embryos, such that the number is shown in the respective bars in panel (g). The asterisks indicate significant differences at P < 0.05*, P < 0.01** or P < 0.001*** using the Mann–Whitney U-test. Ant., anterior; D, dorsal; NS, not significant; Pos. posterior; V, ventral.
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