Fig. 3

Neutrophilic inflammation following tail-fin injury of pik3cd^E1017K/+ embryos and pik3cd^E525K/+ embryos. (A) Composite brightfield and maximal projection of widefield fluorescent (shown in green) micrograph stack, illustrating the TgBAC(mpx:gfp)i114 3 dpf embryo tail-fin injury used to analyse inflammation response. Transection is through the tip of the notochord, and GFP marked neutrophils counted proximal to the circulatory loop. Scale bar: 100 µm. (B) Chart of the tail-fin injury inflammation time course for 72, 3 dpf embryos from a pik3cd^E1017K/+ outcross. Successive neutrophil counts at the injury site are shown as the mean±s.e.m. for embryos subsequently genotyped as pik3cd^E1017K/+ (red) or pik3cd^+/+ (black). Multiple Mann–Whitney tests. (C) Chart with the same data as in B but showing the neutrophil counts for the individual embryos at the 24 h post injury (hpi) end point of the analysis. Mann–Whitney test. Data shown are from one experiment. A repeat experiment with 71 embryos showed the same effect (P<0.0001 at 24 hpi), as did a 36-embryo, 4 hpi time point, pilot experiment (P=0.001). (D) Chart of the tail-fin injury inflammation time course for 71, 3 dpf embryos from a pik3cd^E525K/+ outcross. Successive neutrophil counts at the injury site are shown as the mean±s.e.m. for embryos subsequently genotyped as pik3cd^E525K/+ (red) or pik3cd^+/+ (black). Multiple Mann–Whitney tests. (E) Chart with the same data as in D but showing the neutrophil counts for the individual embryos at the 24 hpi end point of the analysis. Mann–Whitney test. Data shown are from one experiment. A repeat experiment with 96 embryos showed the same effect (P=0.0004 at 24 hpi).