|
Fig. 6 FBXW7 regulates OL MYRF levels in vivo. a Representative images of optic nerves from Fbxw7fl/fl and Plp1-CreERT-Fbxw7fl/fl mice at 1-, 3-, and 6-months post-TAM stained for MYRF. b High-resolution images showing cytoplasmic localization of MYRF in the optic nerve OLs of 6-months post-TAM animals. Arrowheads indicate cytoplasmic localization of MYRF (likely uncleaved precursor) in the Fbxw7fl/fl control. c Representative images of cerebellums from Fbxw7fl/fl and Plp1-CreERT-Fbxw7fl/fl mice at 6-months post-TAM stained for MYRF. d Quantification of nuclear MYRF intensity in Plp1-CreERT-Fbxw7fl/fl (1 m: N = 3, 3 m: N = 3, 6 m: N = 3) optic nerves normalized to controls (1 m: N = 3, 3 m: N = 3, 6 m: N = 4) at 1-, 3-, and 6-months post-TAM. Data shown as mean ± SEM. Statistical significance determined by two-way ANOVA with Tukey’s multiple comparisons test. e Ratio of cytoplasmic relative to total (cytoplasmic and nuclear) MYRF at 6-months post-TAM in Fbxw7fl/fl (N = 4) and Plp1-CreERT-Fbxw7fl/fl mice (N = 3). Data shown as mean ± SEM, statistical significance determined by unpaired, two-tailed Student’s t test. f qRT-PCR quantification of Myrf RNA fold change in the optic nerve from of Fbxw7fl/fl (1 m: N = 3, 12 m: N = 3) and Plp1-CreERT-Fbxw7fl/fl mice (1 m: N = 3, 12 m: N = 6) at 1- and 12- months post-TAM. Data shown as mean ± SEM. Statistical significance determined by two-way ANOVA with Šídák’s multiple comparisons test. g Quantification of nuclear MYRF intensity in Plp1-CreERT-Fbxw7fl/fl cerebellums normalized to controls at 1-, 3-, and 6-months post-TAM. For both genotypes, 1 m (N = 4), 3 m (N = 4), 6 m (N = 4). Data shown as mean ± SEM. Statistical significance determined by two-way ANOVA with Tukey’s multiple comparisons test. h Quantification of NeuN+ cells wrapped in MBP positive membrane in the granule cell layer of the cerebellum in 6- and 12-month post-TAMed Plp1-CreERT-Fbxw7fl/fl (1 m: N = 4, 12 m: N = 3) and control animals (1 m: N = 4, 12 m: N = 3). Data shown as mean ± SEM. Statistical significance determined by two-way ANOVA with Šídák’s multiple comparisons test. i qRT-PCR quantification of Myrf RNA fold change in the cerebellum from Fbxw7fl/fl (1 m: N = 4, 12 m: N = 5) and Plp1-CreERT-Fbxw7fl/fl (1 m: N = 3, 12 m: N = 5) mice at 1- and 12- months post-TAM. Data shown as mean ± SEM. Statistical significance determined by two-way ANOVA with Tukey’s multiple comparisons test. Created in BioRender. Emery, B. (2024) BioRender.com/o45h010/k49b495.