Neutrophil recruitment to the amputation site is increased in mfap4 mutant fish. (a) The tail fin primordium of 3 dpf wild-type (WT, mfap4+/+) or mfap4∆/∆ mutant larvae was amputated (red line) and subsequently analyzed by whole-mount immunofluorescence with the neutrophil marker mpx (black box). (b) Representative images of various time points post amputation of WT and mfap4∆/∆ fins. White dotted lines outline the amputation site. (c) Graphical representation of mean neutrophil numbers in the imaged area near the amputation site [as shown in (b)] of ten biological replicates per genotype for various time points post amputation. Quantification of neutrophil numbers reveals significantly more neutrophils are recruited to the amputation site in mfap4∆/∆ mutants as compared to WT larvae at most analyzed time points (0 hours post amputation (hpa), P = 0.0021; 0.5 hpa, P = 0.0002; 1 hpa, P < 0.0001; 2 hpa, P = 0.1051; 3 hpa, P < 0.0001; 6 hpa; P = 0.0002; assessed by Mann–Whitney-U test). (d) Quantification of neutrophil numbers in the larval tail near the amputation site at the time points shown in (c) relative to uncut tails. The recruitment of neutrophils is not accelerated in mfap4∆/∆ mutants. Error bars in (b) and (c) represent the standard error of the mean (s.e.m.) of ten biological replicates. Graphs in panels (c) and (d) were generated with Microsoft Excel (for Mac 2011, version 14.7.7). This figure was created with Inkscape software version 0.92 (available at https://www.inkscape.org/).
|
