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. 2015 Oct 14;35(41):14042–14056. doi: 10.1523/JNEUROSCI.2781-15.2015

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Copyright © 2015 the authors 0270-6474/15/3514042-15$15.00/0

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Figure 6. — Reduced Aβ pathology in 3xTg-AD/S6K1^+/− mice. A–D, Representative microphotographs of brain sections from 3xTg-AD and 3xTg-AD/S6K1^+/− mice stained with an anti-Aβ₄₂-specific antibody. C and D are high-magnification images of the boxed areas in A and B, respectively. E, Sandwich ELISA measurements of soluble Aβ₄₂ and Aβ₄₀ levels. The levels of both peptides were significantly lower in mice lacking one copy of the S6K1 gene (Aβ₄₀: t₍₁₄₎ = 2.31, p = 0.03; Aβ₄₂: t₍₁₄₎ = 2.81, p = 0.01; n = 8 mice/genotype). F, Sandwich ELISA measurements of insoluble Aβ₄₂ and Aβ₄₀ levels. While the levels of insoluble Aβ₄₀ were similar between the two groups (t₍₁₄₎ 0.92, p > 0.05), the levels of insoluble Aβ₄₂ were significantly lower in 3xTg-AD/S6K1^+/− mice compared with those in 3xTg-AD mice (t₍₁₄₎ = 2.46, p = 0.02; n = 8 mice/genotype). ELISA data were analyzed by Student's t test. G, Representative Western blots of protein extracted from NonTg, 3xTg-AD, 3xTg-AD/S6K1^+/−, and S6K1^+/− mice. To identify full-length APP, blots were probed with 6E10. To identify C99 and C83, blots were probed with CT20, a C-terminal anti-APP antibody. H–J, Quantitative analyses of the blots obtained by normalizing the quantity of a specific protein with its loading control, β-actin. Data were analyzed by one-way ANOVA followed by a Bonferroni's multiple-comparison test (n = 8 mice/genotype; for APP: p < 0.0001, F_(3,28) = 49.05). Post hoc analysis indicated that APP levels were significantly higher in 3xTg-AD and 3xTg-AD/S6K1^+/− compared with the other two groups. No differences were observed between 3xTg-AD and 3xTg-AD/S6K1^+/− mice and between NonTg and S6K1^+/− mice. For C99: p < 0.0001; F_(3,28) = 28.44. Post hoc tests indicated that C99 levels were significantly higher in 3xTg-AD mice compared with the other three groups. C99 levels were also higher in 3xTg-AD/S6K1^+/− compared with those in S6K1^+/− mice. No significant difference was found between NonTg and 3xTg-AD/S6K1^+/− mice and between NonTg and S6K1^+/− mice. For C83: p < 0.0001, F_(3,28) = 15.90. Post hoc tests indicated that C83 levels were significantly higher in 3xTg-AD and 3xTg-AD/S6K1^+/− compared with the other two groups. No differences were observed between 3xTg-AD and 3xTg-AD/S6K1^+/− mice and between NonTg and S6K1^+/− mice. Error bars represent mean ± SEM.