Fig. S7

Cxcl8 mutant sequence, total neutrophils and macrophage recruitment, related to Figure 6

(A) Amino acid sequence of Cxcl8a TALEN-generated mutation. Red arrow indicates mutated residue. Cxcl8a-/- has a 2 a.a. deletion. (B) Representative images of whole Cxcl8a+/- and Cxcl8a-/- sibling larvae at 3 dpf expressing Tg(mpx:gfp) to mark neutrophils. (C) Quantification of total neutrophil number is not significantly different in Cxcl8a-/- compared to heterozygous siblings (n=34, 20). (D) L-plastin antibody staining was performed on Cxcl8a-/- and +/- embryos expressing Tg(mpx:Dendra) at 1, 6, and 20 hpw to label neutrophils (red and green double positive) and macrophages (red only) at the wound. (E) Macrophage numbers at the wound at each time point was not significantly different in Cxcl8a mutants while neutrophil number at the wound (F) was significantly higher at 6 hpw as shown previously (Figure 6B-C, n= 1hpw 16, 10; 6 hpw 12, 12; 20 hpw 14, 5; 1 representative experiment). (G) L-plastin antibody staining was performed on Cxcr2-/- and WT embryos expressing Tg(mpx:Dendra) at 1, 6, and 20 hpw. (H) Macrophage recruitment to the wound was significantly higher in Cxcr2-/- at 6 hpw as was neutrophil presence (I, n= 1hpw 18, 20; 6hpw 23, 25; 20 hpw 20, 21; 1 representative experiment), *p<0.05, n.s.= not significant.