Journal: PLoS ONE

Article Title: Peptides of Presenilin-1 Bind the Amyloid Precursor Protein Ectodomain and Offer a Novel and Specific Therapeutic Approach to Reduce ß-Amyloid in Alzheimer’s Disease

doi: 10.1371/journal.pone.0122451

Figure Lengend Snippet: mThy1-hAPP mice were infused with either PBS or peptides for two weeks, followed by two weeks of rest, using osmotic minipumps. Fixed brains were vibratomed at 40 μm, immunolabeled with MAb against Aβ1–16, followed by FITC-conjugated anti-mouse IgG, and imaged with a scanning confocal microscope. Top Panel: Representative low-power (15x) photomicrographs of vibritome sections (bar, 250 μm). Bottom panel: High power (400x) photomicrographs of amyloid plaques in top panel (bar, 40 μm). Peptides P4 (panels b and h) and P8 (panels c and i) produced marked reductions in Aβ labeling compared to PBS controls (panels a and g).

Article Snippet: For analysis of Aβ deposits, blind-coded 40 μm vibratome sections were immunolabeled as previously described [ ] with mouse monoclonal antibodies (MAb) against human Aβ (clone 82E1, prepared against Aβ 1–16) followed by incubation with fluorescein isothiocyanate (FITC)-conjugated anti-mouse IgG (Vector Laboratories).

Techniques: Immunolabeling, Microscopy, Produced, Labeling

Journal: PLoS ONE

Article Title: Peptides of Presenilin-1 Bind the Amyloid Precursor Protein Ectodomain and Offer a Novel and Specific Therapeutic Approach to Reduce ß-Amyloid in Alzheimer’s Disease

doi: 10.1371/journal.pone.0122451

Figure Lengend Snippet: A. Aβ levels in sections of mouse neocortex that was immunolabeled with a MAb against human Aβ1–16 were quantified as in . Data are expressed as mean ±s.e.m. n = 5. ** p <0.005. B. BACE-1 activity in extracts of neocortex of mice treated with peptides P4, P8 and PBS was determined using the SensiZyme BACE1 activity assay kit. Absorbance was monitored at 405 nm. Data are expressed as mean ±s.e.m. of active BACE-1 in ng/ml n = 5. C. Extracts of neocortex of peptide-treated mice were Western-blotted with primary rabbit Ab against NICD followed by HRP-conjugated goat anti-rabbit IgG. Immunoreactive bands were detected by ECL and the signal intensity of the protein bands was quantified. Top: data are expressed as mean ±s.e.m. n = 5. Bottom: The individual Western blot gel images showing immunoreactive bands. D. After NICD detection, the nitrocellulose membranes were stripped and re-probed with a MAb against APP, followed by HRP-conjugated goat anti-mouse IgG. The signal intensity of the protein bands was then quantified. Top: data are expressed as mean ±s.e.m. n = 5. Bottom: The individual Western blot gel images showing immunoreactive bands.

Article Snippet: For analysis of Aβ deposits, blind-coded 40 μm vibratome sections were immunolabeled as previously described [ ] with mouse monoclonal antibodies (MAb) against human Aβ (clone 82E1, prepared against Aβ 1–16) followed by incubation with fluorescein isothiocyanate (FITC)-conjugated anti-mouse IgG (Vector Laboratories).

Techniques: Immunolabeling, Activity Assay, Western Blot

Journal: Molecular Neurodegeneration

Article Title: Increased amyloidogenic processing of transgenic human APP in X11-like deficient mouse brain

doi: 10.1186/1750-1326-5-35

Figure Lengend Snippet: Effect of X11s in the generation of Aβ in the brains of several transgenic and knock-out mouse lines

Article Snippet: Mouse monoclonal antibodies to human Aβ 82E1 (IBL) and 6E10 (Signet COVANCE), human APP 10D1 (IBL), sAPPα 2B3(IBL), sAPPβswe 6A1 (IBL), actin (Chemicon), flotillin-1 (BD Transduction Laboratories), α-tubulin (Zymed and Santa Cruz Biotechnologies), and X11L/Mint2 (BD Transduction Laboratories) were purchased.

Techniques: Transgenic Assay, Knock-Out

Journal: Molecular Neurodegeneration

Article Title: Increased amyloidogenic processing of transgenic human APP in X11-like deficient mouse brain

doi: 10.1186/1750-1326-5-35

Figure Lengend Snippet: Expression of APP and generation of primary amyloidogenic fragment CTFβ in APP23 mice in the presence or absence of X11L . ( A ) Expression of human APP751swe and detection of human CTFβ (C99) in APP23 mouse brain samples in the presence or absence of X11L and in wild-type mice. Membrane (P100 for APP, CTFβ and flotillin-1) and cytoplasmic (S100 for X11L) fractions of brain regions composed of the cerebral cortex, hippocampus and olfactory bulb from wild-type (WT), APP23 and APP23/X11L-Ko mice (5-6 months old) were analyzed for the expression of transgenic human APP751swe (anti-APP 10D1 antibody), total APP (human APP751swe combined with mouse endogenous APP695, anti-APP/c 8717 antibody), human APP C99 (CTFβ, anti-human Aβ 82E1 antibody), X11L and flotilin-1 by immunoblotting. For human APP, the upper band indicates mature APP751swe (mAPP) and the lower band indicates immature APP751swe (imAPP). For mouse APP (see lanes of WT), the upper two bands are mAPP695 and lower band is imAPP695. To identify CTF species with or without phosphorylation at Thr668, the lysates were subjected to dephosphorylation prior to detection as described in Materials and Methods (for identification of APP and CTF molecules, reviewed in Suzuki and Nakaya ). ( B ) The densities of human APP751swe (left) and C99 (right) bands were standardized to the density of flotillin-1, which was compared with the ratios for APP23 mice that were assigned a reference value of 1.0 (values represent the means ± S.E.). The data were analyzed by Student's t test (n = 10; *, p < 0.05).

Article Snippet: Mouse monoclonal antibodies to human Aβ 82E1 (IBL) and 6E10 (Signet COVANCE), human APP 10D1 (IBL), sAPPα 2B3(IBL), sAPPβswe 6A1 (IBL), actin (Chemicon), flotillin-1 (BD Transduction Laboratories), α-tubulin (Zymed and Santa Cruz Biotechnologies), and X11L/Mint2 (BD Transduction Laboratories) were purchased.

Techniques: Expressing, Transgenic Assay, Western Blot, De-Phosphorylation Assay

Journal: Molecular Neurodegeneration

Article Title: Increased amyloidogenic processing of transgenic human APP in X11-like deficient mouse brain

doi: 10.1186/1750-1326-5-35

Figure Lengend Snippet: Human Aβ levels in APP23 mouse brain samples in the presence or absence of X11L . Quantification of human Aβ40 (left) and Aβ42 (right) in brain samples composed of the cerebral cortex, hippocampus and olfactory bulb from APP23 and APP23/X11L-Ko mice (5-6 months old) was performed by sELISA. Concentrations were normalized to tissue weight. The data were analyzed by Student's t test. Statistical significance is indicated with asterisks (n = 10; *, p < 0.05). The error bars are S.E.

Article Snippet: Mouse monoclonal antibodies to human Aβ 82E1 (IBL) and 6E10 (Signet COVANCE), human APP 10D1 (IBL), sAPPα 2B3(IBL), sAPPβswe 6A1 (IBL), actin (Chemicon), flotillin-1 (BD Transduction Laboratories), α-tubulin (Zymed and Santa Cruz Biotechnologies), and X11L/Mint2 (BD Transduction Laboratories) were purchased.

Techniques:

Journal: Molecular Neurodegeneration

Article Title: Increased amyloidogenic processing of transgenic human APP in X11-like deficient mouse brain

doi: 10.1186/1750-1326-5-35

Figure Lengend Snippet: Human sAPP levels in APP23 mouse brain samples in the presence or absence of X11L . (A) Detection of total human sAPP, sAPPα and sAPPβswe along with human APP, X11L and α-tubulin in APP23 mouse brain tissue in the presence or absence of X11L. Membrane (P100 for APP) and cytoplasmic (S100 for sAPP, sAPPα, sAPPβswe, X11L and α-tubulin) fractions of brain regions composed of cerebral cortex, hippocampus and olfactory bulb from wild-type (WT), APP23 and APP23/X11L-Ko mice (5-6 months old) were analyzed for total human sAPP (sAPPα plus sAPPβswe by 10D1 antibody), human sAPPα (2B3 antibody), human sAPPβswe (6A1 antibody), human APP (10D1 antibody), X11L and α-tubulin by immunoblotting. ( B ) The densities of total human sAPP (left), sAPPα (middle) and sAPPβswe (right) bands were standardized to the density of α-tubulin, which was compared with the ratios for APP23 mice that were assigned a reference value of 1.0 (values represent the means ± S.E.). The data were analyzed by Student's t test (n = 10; **, p < 0.01).

Article Snippet: Mouse monoclonal antibodies to human Aβ 82E1 (IBL) and 6E10 (Signet COVANCE), human APP 10D1 (IBL), sAPPα 2B3(IBL), sAPPβswe 6A1 (IBL), actin (Chemicon), flotillin-1 (BD Transduction Laboratories), α-tubulin (Zymed and Santa Cruz Biotechnologies), and X11L/Mint2 (BD Transduction Laboratories) were purchased.

Techniques: Western Blot

Journal: Molecular Neurodegeneration

Article Title: Increased amyloidogenic processing of transgenic human APP in X11-like deficient mouse brain

doi: 10.1186/1750-1326-5-35

Figure Lengend Snippet: Quantification of amyloid plaques in APP23 mouse brain in the presence or absence of X11L . Immunostaining of coronal sections of the brain region including the cerebral cortex and hippocampus of APP23 (Fig. 4A, upper panels) and APP23/X11L-Ko mice (Fig. 4B, lower panels) at 12 months of age is shown. The brain sections were stained with anti-Aβ 82E1 antibody to detect human Aβ. Stains indicate amyloid plaque. Bar, 300 μm. The data were analyzed by Student's t test (n = 4 serial sections × 6 individuals respectively; **, p < 0.01). The error bars are S.E.

Article Snippet: Mouse monoclonal antibodies to human Aβ 82E1 (IBL) and 6E10 (Signet COVANCE), human APP 10D1 (IBL), sAPPα 2B3(IBL), sAPPβswe 6A1 (IBL), actin (Chemicon), flotillin-1 (BD Transduction Laboratories), α-tubulin (Zymed and Santa Cruz Biotechnologies), and X11L/Mint2 (BD Transduction Laboratories) were purchased.

Techniques: Immunostaining, Staining

Journal: Molecular Neurodegeneration

Article Title: Increased amyloidogenic processing of transgenic human APP in X11-like deficient mouse brain

doi: 10.1186/1750-1326-5-35

Figure Lengend Snippet: Expression of APP in APP-ibl mouse brain in the presence or absence of X11L . ( A ) Expression of human APP695swe and mouse endogenous APP695. Brain regions composed of the cerebral cortex, hippocampus and olfactory bulb from wild-type (WT), APP-ibl and APP-ibl/X11L-Ko mice (4 months old) were homogenized in 10 mM Tris-HCl [pH 7.9] containing 1% (w/v) SDS, 4 M urea, complete protease inhibitor cocktail (Roche Diagnostics), and 1 μM pepstatin. The lysates were analyzed by immunoblotting with anti-human APP (10D1), anti-APP (8717), anti-X11L and anti-α-tubulin antibodies. ( B ) The densities of human APP695swe (left) and human plus mouse APP (right) bands were standardized to the density of α-tubulin, which was compared with the ratios for APP-ibl mice (left) or wild-type mice (right), which were assigned a reference value of 1.0 (values represent means ± S.E.). The data were analyzed by Student's t test (n = 3).

Article Snippet: Mouse monoclonal antibodies to human Aβ 82E1 (IBL) and 6E10 (Signet COVANCE), human APP 10D1 (IBL), sAPPα 2B3(IBL), sAPPβswe 6A1 (IBL), actin (Chemicon), flotillin-1 (BD Transduction Laboratories), α-tubulin (Zymed and Santa Cruz Biotechnologies), and X11L/Mint2 (BD Transduction Laboratories) were purchased.

Techniques: Expressing, Protease Inhibitor, Western Blot

Journal: Molecular Neurodegeneration

Article Title: Increased amyloidogenic processing of transgenic human APP in X11-like deficient mouse brain

doi: 10.1186/1750-1326-5-35

Figure Lengend Snippet: Generation of primary amyloidogenic fragment CTFβ in APP-ibl mice in the presence or absence of X11L . ( A ) Generation of human CTFβ in APP-ibl mice in the presence or absence of X11L. Brain regions composed of the cerebral cortex, hippocampus and olfactory bulb from APP-ibl and APP-ibl/X11L-Ko mice (4 months old) were homogenized in 10 mM Tris-HCl [pH 7.9] containing 1% (w/v) SDS, 4 M urea, complete protease inhibitor cocktail (Roche Diagnostics), and 1 μM pepstatin. The lysates were dephosphorylated as described [ , ] and analyzed by immunoblotting with anti-human Aβ (82E1) and anti-α-tubulin antibodies. ( B ) The density of human CTFβ band was standardized to the density of α-tubulin, which was compared with the ratios for APP-ibl mice that were assigned a reference value of 1.0 (values represent the means ± S.E.). The data were analyzed by Student's t test (n = 10; **, p < 0.01). ( C ) Generation of CTFα and CTFβ in APP-ibl mice in the presence or absence of X11L. Brain lysates of the cerebral cortex, hippocampus and olfactory bulb from wild-type (WT), APP-ibl and APP-ibl/X11L-Ko mice (4 months old) were dephosphorylated and analyzed by immunoblotting with anti-APP/c 8717 and anti-α-tubulin antibodies. ( D ) The densities of C99 (CTFβ), C89 (CTFβ) and C83 (CTFα) bands were standardized to the density of α-tubulin, which was compared with the ratios for APP-ibl mice that were assigned a reference value of 1.0 (values represent means ± S.E.). The data were analyzed by Student's t test (n = 10; **, p < 0.01).

Article Snippet: Mouse monoclonal antibodies to human Aβ 82E1 (IBL) and 6E10 (Signet COVANCE), human APP 10D1 (IBL), sAPPα 2B3(IBL), sAPPβswe 6A1 (IBL), actin (Chemicon), flotillin-1 (BD Transduction Laboratories), α-tubulin (Zymed and Santa Cruz Biotechnologies), and X11L/Mint2 (BD Transduction Laboratories) were purchased.

Techniques: Protease Inhibitor, Western Blot

Journal: Molecular Neurodegeneration

Article Title: Increased amyloidogenic processing of transgenic human APP in X11-like deficient mouse brain

doi: 10.1186/1750-1326-5-35

Figure Lengend Snippet: Human Aβ levels in APP-ibl mouse brain in the presence or absence of X11L . Quantification of human Aβ40 (left) and Aβ42 (right) in the cerebral cortex, hippocampus and olfactory bulb from APP-ibl and APP-ibl/X11L-Ko mice (5-6 months old) was performed by sELISA, with concentrations normalized to tissue weight. The data were analyzed by Student's t test. Statistical significance is indicated with asterisks (n = 10; **, p < 0.01). The error bars are S.E.

Article Snippet: Mouse monoclonal antibodies to human Aβ 82E1 (IBL) and 6E10 (Signet COVANCE), human APP 10D1 (IBL), sAPPα 2B3(IBL), sAPPβswe 6A1 (IBL), actin (Chemicon), flotillin-1 (BD Transduction Laboratories), α-tubulin (Zymed and Santa Cruz Biotechnologies), and X11L/Mint2 (BD Transduction Laboratories) were purchased.

Techniques: