Morpholino-mediated depletion of MYDGF phenocopies neutrophil accumulation at sterile injury observed in mydgf homozygous mutant. (A) Quantification of the tail length of unwounded WT and mydgf^−/− larvae at 6 dpf; two independent replicates with n = 13 +/+ and 10 −/−. (B) Schematic of zebrafish mydgf gene, with sequences targeted by splice-blocking morpholinos #1 and #2 highlighted at the junction of exon 3 and intron 3 and the junction of exon 5 and intron 5, respectively; scale bar = 100 bp. (C) Representative Western blot for zebrafish MYDGF and β-actin from pooled, 2 dpf larvae treated with either mismatch control mo or mo targeting mydgf. (D) Quantification of Western blot in C; representative of three independent replicates. (E–G) Representative brightfield images of Sudan Black staining (E) and quantification of the number of neutrophils in the wound microenvironment of larvae treated with either mismatch control mo and mydgf-targeting mo #1 (F) or mo #2 (G) at 1 and 6 hpw following tail transection of the caudal fin; three independent replicates with n = 52 control and 52 mo #1 at 1 hpw and 53 control and 34 mo #1 at 6 hpw (F), and three independent replicates with n = 47 control and 51 mo #2 at 1 hpw and 44 control and 58 mo #2 at 6 hpw (G); scale bar = 100 µm. (H) Quantification of the total number of neutrophils in 3 dpf whole larvae treated with mismatch control or mo #1 targeting mgdyf; three independent replicates with n = 59 control and 57 mo #1. For F–H, data are expressed as mean with 95% CI; each symbol represents one larva, and different colors represent the results of three independent replicates. ***, P < 0.001; ****, P < 0.0001. P values were calculated by ANOVA with Tukey’s multiple comparisons.