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Fig. 5 Inflammation regulates MG Jak1–Stat3 signaling in zebrafish retinas subjected to stab injury.(A) Violin plot showing the expression of Jak1–Stat3 signaling components in MG and rod progenitors. (B) BEAM analysis of Jak1–Stat3 signaling components (blue) and other markers during MG status transition. (C) Jak1–Stat3 signaling component expression levels in MG at the four time points tested. (D) qPCR analysis of the expression of socs3b, stat3, and inflammatory cytokines in uninjured and stab-injured zebrafish retinas. (E) qPCR analysis of Jak1–Stat3 signaling component expression in stab-injured retinas and were treated with PBS or Dex compared with uninjured retinas (0 days). (F) FISH analysis showing co-localization (arrows) of socs3b mRNA with GFP in the retinas of Tg(gfap:GFP) transgenic fish. (G) FISH showing socs3b mRNA expression in PBS- or Dex-treated retinas at 12 hpi. (H) Zym injection into uninjured retinas induced expression of Jak1–Stat3 signaling components, as assessed by qPCR. (I) Dex treatment suppressed the expression of Stat3-activating cytokines in the injured retina, as assessed by qPCR. (J, K) Microglial ablation by MTZ treatment of Tg(mpeg1:nsfB-mCherry) fish significantly reduced Jak1–Stat3 signaling component expression in the retina. (L) Quantification of proliferating INL cells at 3 dpi in retinas that received the indicated treatments. White asterisks in G and J indicate the site of the stab injury. Data are expressed as mean ± SEM (n = 3).*P < 0.05, **P < 0.01, vs. PBS or control group (one-way analysis of variance followed by Tukey’s post hoc test). BEAM: Branched Expression Analysis Modeling; Dex: dexamethasone; dpi: day post-injury; FISH: fluorescence in situ hybridization; GCL: ganglion cell layer; hpi: hours post-injury; INL: inner nuclear layer; MG: Müller glia; MTZ: metronidazole; ONL: outer nuclear layer; PBS: phosphate-buffered saline; qPCR: quantitative polymerase chain reaction; Zym: zymosan A.