Depletion or inhibition of 12-LOX impairs macrophage migration in zebrafish and mice. (A) Control and alox12 MO-treated Tg(mpeg:GFP) zebrafish underwent tailfin injury at 3 dpf. Representative images of injured zebrafish tails stained with GFP (macrophages, green) and TO-PRO3 (nuclei, red) are shown on the left and quantitation of relative number of migrating macrophages is shown on the right (n = 22–24 fish per condition; *P < 0.05 by unpaired 2-tailed t test). (B) Tg(mpeg:eGFP) zebrafish were treated with vehicle or 10 μM ML355 and then underwent tailfin injury at 3 dpf. Representative images of injured zebrafish tails stained with GFP (macrophages, green) and TO-PRO3 (nuclei, red) are shown on the left and quantitation of relative number of migrating macrophages is shown on the right (n = 7–9 fish per condition; *P < 0.05 by unpaired 2-tailed t test). (C) Description of the chemotaxis assay in vitro using WT and Alox15–/– mouse peritoneal macrophages. (D) Representative images of the porous membrane showing migrating methylene blue–stained macrophages (red arrows) is shown on the left, and quantitation of the number of migrating macrophages is shown on the right (n = 5–7 independent experiments; *P < 0.05 by unpaired 2-tailed t test). Scale bar: 50 μm. All data are presented as mean ± SEM. 12-LOX, 12-lipoxygenase; Tg(mpeg:eGFP), transgenic fish containing enhanced GFP–labeled macrophages; GFP, green fluorescent protein.
|
