Reduced p-MAPK in regenerating caudal fin folds of Shp2-deficient embryos. At 2 dpf, the caudal fin folds of embryos from a ptpn11a+/− ptpn11b−/− incross were amputated and allowed to regenerate. The embryos were fixed at 2 dpa (4 dpf, 2 dpa) and subjected to whole-mount immunohistochemistry using a p-MAPK-specific antibody (Thr202/Tyr204) (green). The embryos were counterstained with DAPI (blue). Maximum-intensity projection images were taken of the caudal fin folds, and all the embryos were genotyped. (A) Representative images of amputated embryo caudal fin folds, with the edges of the fin folds indicated with dashed lines. The number of embryos showing similar patterns/total number of embryos analyzed are indicated in the right-hand column. Scale bars, 100 μm. (B) p-MAPK was quantified by the mean intensity of the region between the notochord and the edge of the caudal fin fold. Equivalent uncut controls were also quantified, and the mean values of all the caudal fin folds are depicted. The statistical significance of the means was determined relative to ptpn11a+/+ ptpn11b−/− zebrafish embryos within the amputated group, and likewise within the uncut group. **, P < 0.01; *, P < 0.05; the error bars represent standard deviations.
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