The expression level of the nomo1 mRNA during development and generation of nomo1 mutant zebrafish. (A) Relative nomo1 transcript levels at different developmental stages (biological replicates, n = 3). (B) Diagram of zebrafish nomo1 gene and mutation induced by CRISPR/Cas9, gray boxes indicate exons. Target site was on Exon 7 resulting in a 1-base deletion (shown in sequence). The mutation results in early termination of translation after exon 7, before the functional domain of Nomo1, EMC7-beta-sandw. (C) Reduced expression of nomo1 mRNA in the whole embryo at 48 hpf, heads at 14 dpf and brain tissues at 2 mpf of nomo1−/− zebrafish analyzed using RT-qPCR (biological replicates, n = 4). (D) Brain weight of nomo1 + /+ and nomo1−/− shows significant differences (biological replicates, n = 6, 20 brains for each replicate group). Scale bar = 1 mm. (E) Quantitative analysis of brain cells or tissues (biological replicates, n = 6). (F) Brains of nomo1−/− exhibit relatively fragile brain tissue, arrows indicate tissue that collapses immediately after sectioning due to being too fragile. Red dashed circles indicate nucleus with abnormal neuronal development. Scale bar = 25 μm. Data information: Data are analyzed using unpaired t test and presented as the means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Source data are available online for this figure.
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