Journal: Molecular Neurodegeneration

Article Title: Anti-malaria drug artesunate prevents development of amyloid-β pathology in mice by upregulating PICALM at the blood-brain barrier

doi: 10.1186/s13024-023-00597-5

Figure Lengend Snippet: Primary and secondary antibodies used for Immunoblotting

Article Snippet: Rabbit anti-human carboxy terminus of APP, cross-reacts with mouse APP and APP-CTFs (Cell Signaling, 76600S, 1:1000) , HRP-conjugated donkey anti-rabbit (Invitrogen, A16023,1:3000).

Techniques: Sequencing

Journal: Molecular Neurodegeneration

Article Title: Anti-malaria drug artesunate prevents development of amyloid-β pathology in mice by upregulating PICALM at the blood-brain barrier

doi: 10.1186/s13024-023-00597-5

Figure Lengend Snippet: Primary and secondary antibodies used for Immunoblotting

Article Snippet: Rabbit anti-human carboxy terminus of APP, cross-reacts with mouse APP and APP-CTFs (Cell Signaling, 76600S, 1:1000) , HRP-conjugated donkey anti-rabbit (Invitrogen, A16023,1:3000).

Techniques: Sequencing

Journal: Molecular Neurodegeneration

Article Title: Anti-malaria drug artesunate prevents development of amyloid-β pathology in mice by upregulating PICALM at the blood-brain barrier

doi: 10.1186/s13024-023-00597-5

Figure Lengend Snippet: Primary and secondary antibodies used for Immunoblotting

Article Snippet: Rabbit anti-human carboxy terminus of APP, cross-reacts with mouse APP and APP-CTFs (Cell Signaling, 76600S, 1:1000) , HRP-conjugated donkey anti-rabbit (Invitrogen, A16023,1:3000).

Techniques: Sequencing

Journal: Journal of Neuroinflammation

Article Title: Amyloid precursor protein modulates β-catenin degradation

doi: 10.1186/1742-2094-4-29

Figure Lengend Snippet: APP expression induced degradation of β-catenin . A. Overexpression of APP reduces β-catenin levels. Human APP 695 or the control LacZ was expressed in primary neurons via HSV vectors at 1 or 2 IU per cell during 14 h of infection. The control HSV vector carries the β-galactosidase gene (LacZ). B. Quantitative densitometric analyses of western blots from 4 independent experiments. An average 44% reduction in normalized mean optical density (representing protein levels) was observed for 2 IU/cell of APP compared to the corresponding LacZ control ( p < 0.05, t -Test). Shorter exposures were used for densitometry analyses; longer exposures also showed the expression of endogenous APP, but densitometric analyses were not possible due to the solidified chemiluminescence reaction products.

Article Snippet: The following antibodies were used: rabbit anti-β-catenin and mouse anti-γ-tubulin (both from Sigma); anti-phosphorylated β-catenin antibodies, S33/37/T41 and S45 (both from Cell Signaling); rabbit anti-APP antibody (369, a gift from Dr. Sam Gandy).

Techniques: Expressing, Over Expression, Infection, Plasmid Preparation, Western Blot

Journal: Journal of Neuroinflammation

Article Title: Amyloid precursor protein modulates β-catenin degradation

doi: 10.1186/1742-2094-4-29

Figure Lengend Snippet: APP facilitates β-catenin degradation by increasing β-catenin phosphorylation at S33/37/T41 residues . A. Primary neurons infected with APP contained less steady-state β-catenin and higher levels of phosphorylated S33/37/T41 than did neurons infected with control LacZ (2 IU virus per cell). Levels of phosphorylated S45 did not appear to change. B. Quantitative analyses of western blots from 3 independent experiments. APP significantly increased β-catenin phosphorylation at S33/37/T41 residues (p = 0.02, t -Test), but not at S45 residue (p = 0.19, t -Test).

Article Snippet: The following antibodies were used: rabbit anti-β-catenin and mouse anti-γ-tubulin (both from Sigma); anti-phosphorylated β-catenin antibodies, S33/37/T41 and S45 (both from Cell Signaling); rabbit anti-APP antibody (369, a gift from Dr. Sam Gandy).

Techniques: Infection, Western Blot

Journal: Journal of Neuroinflammation

Article Title: Amyloid precursor protein modulates β-catenin degradation

doi: 10.1186/1742-2094-4-29

Figure Lengend Snippet: Suppression of endogenous APP results in increased total β-catenin and decreased S33/37/T41-phosphorylated β-catenin . Primary neurons were infected with APPshRNA (APP1996) or missense shRNA viruses (0.5 IU/cell) for 14 h before harvest and protein analyses. The mean optical densities of 3 independent experiments for each protein analyzed are presented in the Y-axis in the bar graphs. The statistical significance of difference is also presented in each bar graph ( p values from t -Test) for each protein analyzed (labelled on the Y-axis). Representative blots are shown on the right. Loading controls were γ-tubulin reprobed from the same blots.

Article Snippet: The following antibodies were used: rabbit anti-β-catenin and mouse anti-γ-tubulin (both from Sigma); anti-phosphorylated β-catenin antibodies, S33/37/T41 and S45 (both from Cell Signaling); rabbit anti-APP antibody (369, a gift from Dr. Sam Gandy).

Techniques: Infection, shRNA

Journal: Journal of Neuroinflammation

Article Title: Amyloid precursor protein modulates β-catenin degradation

doi: 10.1186/1742-2094-4-29

Figure Lengend Snippet: APP expression downregulates β-catenin levels in membrane and cytosolic fractions but not in the nuclear fraction . A. β-Catenin levels from membrane and cytosolic fractions isolated from primary neurons. Wild type or FAD APPs were overexpressed in primary neurons via HSV-mediated gene transfer using LacZ as the control. 10 μg protein per sample was analyzed in each fraction. B. Selected z-series of single cells. Scale bar, 2 μm. C. Composite images from z-series. Scale bar, 5 μm. In both B and C, primary neurons were infected with APP or the control LacZ viruses at 2 IU/cell for 14 h. Cells were then fixed and stained with rabbit anti-β-catenin and Alexa fluor 488 goat anti-rabbit. Images were collected at the same settings for fluorescence intensity comparison.

Article Snippet: The following antibodies were used: rabbit anti-β-catenin and mouse anti-γ-tubulin (both from Sigma); anti-phosphorylated β-catenin antibodies, S33/37/T41 and S45 (both from Cell Signaling); rabbit anti-APP antibody (369, a gift from Dr. Sam Gandy).

Techniques: Expressing, Isolation, Infection, Staining, Fluorescence

Journal: Journal of Neuroinflammation

Article Title: Amyloid precursor protein modulates β-catenin degradation

doi: 10.1186/1742-2094-4-29

Figure Lengend Snippet: β-Catenin and cyclin D1 are significantly elevated in hippocampal CA1 pyramidal cells in APP knockout mice compared to those in WT controls . Brain sections from APP knockout and WT mice (n = 4 each) were co-labelled with rabbit anti-β-catenin and mouse anti-cyclin D1 together with Alexa Fluor 594 and Alexa Fluor 488 secondary antibodies. The sections were also counter-stained with DAPI to visualize the cell body layer. Images were captured at the same settings with a Nikon Eclipse 600 microscope. Representative images from an APP knockout and a WT animal are presented in A. The pixel density of specific staining was randomly sampled in the CA1 cell body layer and the mean values from each group of animals are shown in the bar graphs: B. β-catenin, two-tail t-Test, APP knockout vs AT, p = 0.001; C. cyclin D1, two-tail t-Test, APP knockout vs WT, p = 0.02.

Article Snippet: The following antibodies were used: rabbit anti-β-catenin and mouse anti-γ-tubulin (both from Sigma); anti-phosphorylated β-catenin antibodies, S33/37/T41 and S45 (both from Cell Signaling); rabbit anti-APP antibody (369, a gift from Dr. Sam Gandy).

Techniques: Knock-Out, Staining, Microscopy