Figure 1.
Wound-Induced Inflammation Triggers ROS Production and Oxidative Damage
(A–K) Wounding and inflammation (green immune cells, srp > GFP, A–C) in Drosophila embryos are associated with increased ROS (magenta, DHE staining) production (A, schematic; B, pre-wound; C, 1 h post-wound; arrowheads indicate ROS within immune cells), oxidative damage (arrowheads, magenta 8-oxo-dG, D and E; quantified in F), γH2AvD puncta (arrowheads, magenta, G and H; quantified in I), and PARylation (blue, J and K). % 8-oxo-dG and % γH2AvD refer to percent (%) of area measured that is positive for marker of interest after thresholding.
(L–O) Inhibition of wound inflammation (macrophage ablation using srp > reaper [L] and trpm-RNAi [M]) accelerates the rate of wound closure compared to controls (quantified in L and M, n > 20 for each condition). Macrophage ablation is associated with reduced ROS production (magenta DHE staining) before (N) and after (arrowheads, O) wounding compared to controls (B and C).
Wound edge represented by dashed yellow outlines in (C), (E), (H), (K), and (O). Scale bars represent 10 μm in (B)–(E), (G), (H), (J), (K), (N), and (O). Data represented as mean ± SEM; ∗p < 0.05 and ∗∗p < 0.01 via the Mann-Whitney Test (F), one-way ANOVA followed by Dunn’s multiple comparisons test (I), or multiple t tests followed by Holm-Sidak multiple comparisons correction (L and M).