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Figure 4


Fig. 4. The consequences of WASp1- and WASp2-morpholino injections on leukocyte recruitment to a fin wound in 3-dpf larvae. (A-C) Typical 90-minute wounds in control- (A), WASp1-morpholino-injected (B) and WASp2-morpholino-injected (C) larvae after staining with Sudan Black to reveal neutrophil influx to the fin wound. (D-F) Similar wounds, but harvested at 3 hours and immunostained with antibody against L-plastin to reveal the influx of macrophages and neutrophils. (G) Images from a typical fluorescent time-lapse movie of control- and WASp1-morpholino-injected (MO) Tg(lyz:EGFP) fish after fin wounding; the white dotted ovals indicate wound location. See also supplementary material Movies 3 and 4. (H) qPCR analysis of egfp, lysC, wasp1 and wasp2 expression within EGFP-marked myelomonocytic cells of 3-dpf Tg(lyz:EGFP) larvae and Tg(lyz:EGFP) whole kidney marrow, relative to that within EGFP-negative cells. Error bars represent standard deviations. (I) FACScan analysis of EGFP-labelled compartments within wild-type, WASp1- and WASp2-depleted 3-dpf larvae. The y-axis indicates the percentage of green cells from total cells analysed, and reveals no significant difference between control and WASp-morphant fish. Averages and standard deviations calculated from two separate experiments for each sample are shown. Scale bars: A-F, 25 µm; G, 15 µm.





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