MYDGF depletion alters neutrophil motility in the wound microenvironment. (A) Caudal fins of 3 dpf WT or mydgf^−/− zebrafish larvae with mCherry-labeled neutrophils were wounded by tail transection, and fixed at 1 and 6 hpw. Neutrophils were counted at the wound, distal to the tip of the notochord. (A and B) Representative images (A) and quantification (B) of neutrophils in the wound are shown; three independent replicates with n = 27 +/+ and 20 −/− at 1 hpw and 42 +/+ and 37 −/− at 6 hpw; scale bar = 100 µm. (C) Quantification of total number of mCherry-labeled neutrophils in 3 dpf WT and mydgf^−/− whole larvae; three independent replicates with n = 55 WT and 58 −/−. In B and C, data are expressed as mean with 95% CI; each symbol represents one larva, and different colors represent independent replicates. *, P < 0.05; ***, P < 0.001. P values were calculated by ANOVA with Tukey’s multiple comparisons. (D) Representative serial images from time-lapse imaging 0–6 h following tail transection of the caudal fin of WT and mydgf^−/− larvae; see Video 1. Lines represent neutrophil tracks over time, with warmer colors indicating a longer time since track start; four independent replicates with n = 7 +/+ and 11 −/− larvae; scale bar = 100 µm. (E) Quantification of the number of neutrophils at the wound microenvironment over the course of time-lapse imaging. (F) Quantification of the instantaneous speed during the later phase (3–6 hpw) of neutrophil recruitment; 0 min represents the time at which each neutrophil enters the wound microenvironment.