Fig. 3

hspd1 mutants have impaired neutrophil migration towards the injury site. (a) Neutrophil migration is triggered by hair cell ablation. Neutrophils were labelled by Tg(mpx:EGFP) and pictures were taken after 1 h of CuSO₄ treatment. Arrows point to neutrophil accumulation in lateral line neuromast areas. (b) Quantification of the neutrophils that migrated to the regions surrounding the neuromasts; scoring was done before the genotypes were known. The reduction is significant (n=12, P=0.001). (c) Neutrophil migration triggered by caudal fin amputation. Pictures were taken at 17 h post amputation, when the difference in the number of migrated neutrophils between control and mutant embryos was greatest. (d) Quantification of the neutrophils migrated to the amputated fin at 17 h post amputation; scoring was done before the genotypes were known. Red boxes demarcate the areas used for quantification. The reduction is significant (n=10, P<0.001). (e) Neutrophil migration triggered by injection of LPS. The top panels show the control embryos injected with phenol red buffer and BSA. The red box in the top right panel shows a low number of mpx:EGFP cells accumulated in the BSA injection area. The bottom panels show control and hspd1 mutant embryos injected with 150 pg LPS. Red dotted boxes show that LPS injection strongly attracts neutrophils into the injection area. (f) Quantification of neutrophils migrated to the LPS injection site in hspd1 control and mutant embryos. Red dotted boxes demarcate the areas used for quantification. There is no significant difference between mutant and control in neutrophil’s ability to migrate (n=10, P=0.291). Asterisks indicate a significant difference. Bars = 200 μm. BSA, bovine serum albumin; LPS, lipidpolysaccharide.