Figure 1 | Scientific Reports

Figure 1

From: RBFOX3/NeuN is Required for Hippocampal Circuit Balance and Function

Figure 1

Validation and phenotypic characterization of Rbfox3−/− mice.

(a) Schematic of the targeting locus on the Rbfox3 gene. A LacZ-Neo cassette was inserted into the intron between exon 6 and 7 and FRT sites flanked the cassette. LoxP sites flanked exons 7 to 9. (b) Genotyping by PCR analysis of tail genomic DNA [WT (+/+), heterozygous (+/−) and homozygous (−/−) mutant] using Rbfox3, LacZ and Neo primers (indicated in a). (c) Quantitative RT-PCR analysis of cerebral cortical Rbfox3 transcripts from WT and Rbfox3 homozygous knockout (Rbfox3−/−) (KO) mice. Student’s t-test, two-tailed **P < 0.01, n = 4 per group. (d) Western blotting analysis of hippocampal RBFOX3 protein from WT and KO mice probed with antibodies to RBFOX3 and ACTIN. Student’s t-test, two-tailed *P < 0.05, n = 4–6 per group. (e) Hippocampal regions from WT and KO mice immunostained with RBFOX3 antibody (green) and counter-stained with DAPI (blue). Scale bar = 200 μm. (f) Somatosensory cortical regions from WT and KO mice immunostained with anti-RBFOX3 antibody (green) and counter-stained with DAPI (blue). Scale bar = 200 μm. (g) Body weights and (h) brain weights from WT and KO mice. Student’s t-test, two-tailed, *P < 0.05, ***P < 0.001, n = 7-13 per group. Neuronal integrity determined with western blotting analysis of neurofilament protein heavy chain (NF200-H) (i, left) and its phosphorylated (pNF-H) (i, middle) and non-phosphorylated (NF-H) form (i, right), neurofilament protein light chain (j), neurofilament protein medium chain (k) in the hippocampus of WT and KO mice. Student’s t-test, two-tailed or Mann-Whitney rank sum test *P < 0.05, n = 6-8 per group. (l) White matter volume determined with MRI analysis in the whole brain of WT and KO mice. Student’s t-test, two-tailed, *P < 0.05, n = 6 per group.

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