Fig. 2
Fig. 2
a Experimental setup. hpt, hours post-trauma. b, c The expression of il1b?+?and il6?+?cells (red arrow) in zebrafish larvae was measured by WISH at 3 hpt. Scale bar, 200?µm. d qPCR showed that the mRNA levels of various cytokines or chemokines changed in the traumatic site/tail and the brain/head compared to CTRL. Independent t test, *, p?<?0.05, **, p?<?0.01. e WISH showed increased il1b?+?cells (red arrow) in the brain/head, body/trunk, and traumatic site/tail of zebrafish larvae at 24 hpt. Scale bar, 100?µm. h, head. b, body. t, tail. f Design of the transcription activator-like effector nucleases (TALEN) targeting zebrafish il1b. g, h il1b mutation (il1bMut) was confirmed by the restriction fragment length polymorphism (RFLP) assay and Sanger sequencing (?7 bp). (i) qPCR showed that the mRNA levels of various cytokines were decreased in il1bMut zebrafish brains compared to CTRL brains. Independent t test, **, p?<?0.01. j?l il1bMut rescued the increased mpeg1?+?macrophages or coro1a?+?leukocytes in the zebrafish brain at 24 hpt. Two-way ANOVA with Tukey?s HSD post hoc test, **, p?<?0.01 compared to CTRL. Scale bar, 40?µm.