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Fig. S1
Cxcr1 morpholino and mRNA rescue, related to Figure 1
(A) Amino acid sequence of Cxcr1 TALEN-generated mutation. Red arrow indicates mutated residue. (B) Quantification of neutrophils at the wound at 12 and 24 hpw are similar in Cxcr1-/- compared to Cxcr1+/- siblings (n= 12hpw 15,9; 24hpw 22,9). (C) Sudan black staining of neutrophils in Control MO and Cxcr1 MO injected larvae at 1-6 hpw. (D) Quantification of neutrophils at the wound reveals decreased recruitment in Cxcr1 morphants (n= 1hpw 21, 15; 3hpw 24, 14; 6hpw 19,14, ctrl/Cxcr1 MO respectively). (E) Sudan black staining post-wound of Cxcr1 het and mutant larvae injected with cxcr1 mRNA. (F) Quantification of neutrophils at the wound shows cxcr1 mRNA rescues neutrophil recruitment in Cxcr1-/- (n=1hpw 17, 25, 12, 31; 3hpw 17, 24, 11, 26). (G) Representative images of whole Cxcr1+/- and Cxcr1-/- sibling larvae at 3 dpf expressing Tg(mpx:gfp) to mark neutrophils. (H) Representative imaging of the otic vesicle (white dotted outline) 2 hours after injection with supernatant from HEK cells overexpressing zebrafish Cxcl8a or control supernatant. Supernatant was injected into the otic vesicle of 3 dpf Tg(mpx:gfp) WT or Cxcr1-/- larvae (neutrophils in green). (I) Number of neutrophils in the otic vesicle of Cxcl8a or control supernatant injected larvae (n= ctrl 35, 35; Cxcl8 63, 53). (J) Neutrophil fold change response to Cxcl8a over control supernatant. *p<0.05, n.s.=not significant.