Fig 1: In situ hybridisation of rnf38 in tilapia brain. (A) Schematic sagittal diagram of tilapia brain. (B,Bi) In situ hybridisation staining with rnf38 anti-sense RNA probe shows distinct cell-labelling. (Bi) shows a higher magnification of the boxed area in (B). (C,Ci). In situ hybridisation staining with rnf38 sense RNA probe shows no hybridisation signal. Ci shows a higher magnification of the boxed area in (C). Scale bars: (B,C), 200 μm; (Bi,Ci), 50 μm.
Fig 2: Rnf38 mRNA expression in the brain of tilapia. (A) Schematic sagittal diagram of tilapia brain with approximate locations of coronal sections (B–E). (B–E) Representative low magnification photomicrographs of coronal sections are shown on the left and outlined as a mirror image on the right. (Bi,Bii,Ci,Cii,Di,Dii,Ei,Eii) show a higher magnification of the boxed area in (B–E) respectively. Scale bars: (B–E), 500 μm; (Bi,Bii,Ci,Cii,Di,Dii,Ei,Eii), 50 μm. For abbreviations, see Table 1.
Fig 3: Double-labelling immunocytochemistry of RNF38 protein with neuronal marker HuC/D or glial marker GFAP. (A–C) Neuronal marker HuC/D (A, green), RNF38 peptide (B, red), merged image (C). (D–F) Glial marker GFAP (D, green), RNF38 protein (E, red) and merged image (F). Scale bars: (A–F), 10 μm.
Fig 4: Cloned sequence of tilapia rnf38 mRNA. (A) Alignment of cloned rnf38 partial sequence with predicted sequence from GenBank (XM 005467588.3; position 2,490–2,789). (B) Nucleotide similarity between identified tilapia rnf38 partial sequences with astatotilapia (Haplochromis burtoni), medaka (Oryzias latipes), mouse (Mus musculus), rat (Rattus norvegicus), and human (Homo sapiens).
Fig 5: Distribution of rnf38 mRNA in the telencephalon, preoptic area, optic tectum, midbrain, hypothalamus, cerebellum and hindbrain of tilapia. (A) Brain areas collected during microdissection (telencephalon, n = 5; preoptic area, n = 6; optic tectum, n = 8; midbrain, n = 7; hypothalamus, n = 6; cerebellum, n = 5; and hindbrain, n = 5) for rnf38 gene expression study. (B) DNA copy number of rnf38 mRNA in telencephalon, optic tectum, hypothalamus, cerebellum and hindbrain of male tilapia. Data represent the means ± SEM.
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