Fig. 1

Analysis of Aβ-peptide levels and -aggregation in brains of 5xFAD mice. a Hippocampal and cortical brain extracts of 2-, 5-, and 8-month-old AD (n = 5) mice were used for Aβ42 peptide ELISAs. As a control (Ctrl), a pool of wt mouse brain extracts was tested in parallel, which was produced by mixing equal protein amounts of tissue extracts derived from 5 different wt mice per age and tissue. The graph represents the mean ± SD of five biological replicates of tg mice per age and tissue. Statistical analysis: Unpaired, two-tailed t-test between hippocampal and cortical samples of mice of the same age (**, p = 0.0054). b Quantification of Aβ aggregates retained on filter membranes (Additional file 1: Fig. S2a) was performed using the Aida image analysis software. Per tissue and age, extracts of five different mice were analyzed. All data are expressed as mean ± SD of five biological replicates. Statistical significance was assessed between hippocampal and cortical samples of mice with the same age using an unpaired, two-tailed t-test (**, p = 0.0016). c Pearson correlation between Aβ42 peptide levels determined by ELISA (blue line, right axis) and Aβ aggregates determined by MFA (purple line, left axis); hippocampal tissue samples were analyzed. The statistical significance of the association between the Aβ42 peptide levels and Aβ aggregates was measured with a two-tailed t-test (*, p = 0.024). d Plaque load was determined in hippocampus and cortex of 5xFAD mice at different ages (n = 5 mice per age and tissue). Plaques were immunohistochemically stained with the antibody 6E10. Data represent mean ± SD. The statistical significance was assessed between hippocampal and cortical tissues from the same age using an unpaired, two-tailed t-test (*, p = 0.0204; ***, p = 0.0009; ****, p < 0.0001). e Plaque counts were determined in hippocampal and cortical tissues of 2-, 5-, and 8-month-old 5xFAD mice (n = 5 per age and tissue) by antibody 6E10 and ThioS staining. Data represent mean ± SD. The statistical significance was assessed with the antibody 6E10 (*, p = 0.0141; ***, p = 0.0002; ****, p < 0.0001) and ThioS staining (ns; ***, p = 0.0003; ****, p < 0.0001) each for hippocampal and cortical tissues of mice with the same age using an unpaired, two-tailed t-test. f Immunofluorescence analysis of dense core (top row) and diffuse plaques (bottom row) in a brain slice of an 8-month-old 5xFAD mouse. Red indicates 6E10 immunoreactive material. Green indicates fibrillar Aβ material stained with ThioS. Yellow indicates merged signal. g Fractionation of hippocampal and cortical brain extracts derived from 8-month-old 5xFAD mice. Fractions from sucrose density gradient centrifugations of 10,000 × g membrane pellets were analyzed by immunoblotting using antibodies detecting APP and amyloid-β (antibody 6E10), marker proteins of the endoplasmic reticulum (Calnexin), lipid rafts (Flotillin), the Golgi apparatus (Golgin97), lysosomes (Lamp1), and mitochondria (VDAC, NDUFB3). Equal exposure times per antibody for hippocampus and cortex are shown. Per fraction, equal volumes were loaded. From the solubilized pellet (P10,000 × g), 5 µg was loaded