Fig. 2

Fig. 2

Neutrophil deficiency in csf3r null F2 embryos.

(a–d) Fluorescence micrographs of 3 dpf Tg(mpx:EGFP) embryos showing that several representative csf3r mutant null allelotypes (b–d) have substantially reduced numbers of fluorescent neutrophils compared to WT (a). Panel b is a representative homozygous csf3r^3/3 null embryo, and panels c-d show representative compound heterozygous null embryos of the two allelotypes shown. (e) Quantification over 2–5 days post-fertilisation (dpf) of tail region neutrophil numbers in embryos of control (WT) matings and incrosses of 3 different pairs of csf3r null parents of the csf3r allelotypes shown. Data are mean ± SD. Within-day comparisons across genotypes are analysed by unpaired two-tailed t-tests, pooling data from the mutant compound heterozygous allelotypes shown, which are not significantly different to each other. These parental allelotype combinations were randomly selected, being those pairs that laid of those that were set up. They are not intended to be comprehensive, but they do demonstrate a consistent non-complementing neutrophil-depletion phenotype encompassing five embryonic allelotypes (1/2, 1/3, 2/2, 2/3 and 3/3). (f) Embryos carrying single csf3r null alleles have an intermediate neutrophil deficiency. A mix of heterozygous csf3r^wt/(1or2) embryos in a 1:1 ratio were generated by outcrossing a parent of genotype csf3r^1/2 to WT. Their neutrophil numbers at 3 dpf are compared with the non-contemporaneous csf3r^WT/WT and pooled csf3r^−/− mutant 3 dpf groups of panel (e). The mutant data were pooled as none of the mutant allelotype groups is significantly different to any other. p-values are from one-way ANOVA with Tukey’s multiple comparisons test. p < 0.0001. (g) csf3r null embryos have equivalent survival to WT embryos up to 5 days post-fertilisation (dpf). Kaplan-Meier plots compared by Wilcoxon rank sum test.