Figure 3

Copper-stressed macrophages and neutrophils respond more quickly to A. hydrophila infection. (A) Schematic view of tail injury, A. hydrophila infection, and GFP-positive cell counting, with immersion after injury being used in the assays. (B) The recruitment of GFP-positive macrophages and neutrophils around injury locus (in the fixed red boxes) were analyzed at 0, 2, 4, and 6 hpi, using Tg (coro1a:eGFP) transgenic fish. (C–E) Quantification analysis showed a significant increase in the number of GFP-positive cells in the area near the injury locus in the copper-stressed Tg(coro1a:eGFP) larvae compared with the no-copper-stressed control at 0, 2, 4, and 6 hpi (C), in the copper-stressed Tg (mpx:eGFP) larvae (D), and in the copper-stressed Tg (lyz:eGFP) larvae (E). Three biological replicates were performed. Analysis with hypergeometric distribution in R-console software for bi-modal distribution to exhibit individual variations in each group in one experiment. Data are presented as mean ± SD. ***P < 0.001; **P < 0.01; *P < 0.05; and ns, no significance.