Fig. S2

lta4h Deficiencies Do Not Affect Macrophage or Neutrophil Number or Recruitment, Related to Figure 4

(A and B) Live uninfected animals were stained with neutral red at three dpf to quantitate the number of macrophages in control, zm5961 mutant, and lta4h morphant animals.

(A) The number of neutral red positive cells posterior to the yolk tube was determined. No significant differences in macrophage number were observed (one-way ANOVA).

(B) Example of neutral red positive cells (arrows) stained at three dpf in wild-type and lta4h morphant animals. Scale bar, 25 μm.

(C and D) Uninfected fixed animals were stained with Sudan Black at three dpf to quantitate the number of neutrophils present.

(C) Quantitation of the number of neutrophils posterior to the yolk tube. No significant differences among control, zm5961, and lta4h morphant were found (one-way ANOVA).

(D) Example of Sudan Black neutrophil staining in the tail of wild-type and morphant animals. Scale bar, 100 μm.

(E) LTA4H and control morphant animals were injected in the hindbrain ventricle with 150-200 bacteria at 24 hpf. The number of recruited macrophages was assessed 6 hr later by DIC microscopy. No significant differences between control and morphant were found using Student′s unpaired t test.

(F) WT animals compared to zm5961 mutants that had also been injected with the lta4h morpholino. Animals were infected in the hindbrain ventricle and assayed for macrophage recruitment as described above.