human app  (Cell Signaling Technology Inc)

Journal: Receptors (Basel, Switzerland)

Article Title: Gelsolin Facilitates Estrogen Receptor Beta Nuclear Translocation and Transcriptional Repression of Genes Associated with Alzheimer Disease

doi: 10.3390/receptors4020010

Figure Lengend Snippet: ERβ1 represses APP and ITPKB promoter activity. Luciferase activity was measured in SK-N-SH cells following transfection with human promoters for APP-luc ( A ) and ITPKB-luc ( B ) and varying concentrations of ERβ1 ± siRNA-mediated GSN knockdown. ( C–E ) Luciferase was measured in SK-N-SH cells following transfection with human promoters for APP-luc ( C ) and ITPKB-luc ( D ), or HPRT-luc ( E ) and 20 ng ERβ1 ± siRNA-mediated GSN knockdown. Relative promoter activity was calculated based on percentage (%) change from promoter-only group. Data sets are from 3 independent biological replicates with 12 technical replicates each. Data are depicted as mean ± SEM. An ** denotes p < 0.01, ***, p < 0.001, **** p < 0.0001.

Article Snippet: Reporter constructs for human APP (GoClone cat #S719381), human ITPKB (GoClone cat #S701052), human HPRT (GoClone cat #S705060) and an empty promoter-less plasmid (GoClone cat #S790005) were purchased from SwitchGear Genomics, Menlo Park, CA, USA.

Techniques: Activity Assay, Luciferase, Transfection, Knockdown

Journal: Receptors (Basel, Switzerland)

Article Title: Gelsolin Facilitates Estrogen Receptor Beta Nuclear Translocation and Transcriptional Repression of Genes Associated with Alzheimer Disease

doi: 10.3390/receptors4020010

Figure Lengend Snippet: ERβ1 and GSN regulation of APP and ITPKB promoters is independent of E2. Luciferase activity was measured in SK-N-SH cells following transfection with human promoters for APP-luc ( A ) or ITPKB-luc ( B ), plus 20 ng ERβ1, siRNA-GSN and ±10 nM E2. Relative promoter activity was calculated based on % change from promoter-only group. Data sets are from 3 independent biological replicates with 12 technical replicates each. Data are depicted as mean ± SEM. An * denotes p < 0.05, **, p < 0.01, ****, p < 0.0001.

Article Snippet: Reporter constructs for human APP (GoClone cat #S719381), human ITPKB (GoClone cat #S701052), human HPRT (GoClone cat #S705060) and an empty promoter-less plasmid (GoClone cat #S790005) were purchased from SwitchGear Genomics, Menlo Park, CA, USA.

Techniques: Luciferase, Activity Assay, Transfection

Journal: Receptors (Basel, Switzerland)

Article Title: Gelsolin Facilitates Estrogen Receptor Beta Nuclear Translocation and Transcriptional Repression of Genes Associated with Alzheimer Disease

doi: 10.3390/receptors4020010

Figure Lengend Snippet: ERβ1Δ3 represses APP and ITPKB promoter activity. Luciferase activity was measured in SK-N-SH cells following transfection with human promoters for APP-luc ( A ) or ITPKB-luc ( B ), plus varying concentrations ERβ1Δ3 ± siRNA-GSN. Relative promoter activity was calculated based on percentage (%) change from promoter-only group. Data sets are from 3 independent biological replicates with 12 technical replicates each. Data are depicted as mean ± SEM. An * denotes p < 0.05, **, p < 0.01, ****, p < 0.0001.

Article Snippet: Reporter constructs for human APP (GoClone cat #S719381), human ITPKB (GoClone cat #S701052), human HPRT (GoClone cat #S705060) and an empty promoter-less plasmid (GoClone cat #S790005) were purchased from SwitchGear Genomics, Menlo Park, CA, USA.

Techniques: Activity Assay, Luciferase, Transfection

Journal: Brain

Article Title: Tau, synapse loss and gliosis progress in an Alzheimer’s mouse model after amyloid-β immunotherapy

doi: 10.1093/brain/awae345

Figure Lengend Snippet: Amyloid–tau coexpression exacerbates tau tangles and axonal dystrophy in the APP/PS1xTau22 mouse model. ( A ) Amyloid-β (Aβ)-directed antibody reveals diffuse and dense-core Aβ plaques in the hippocampus and cortex of APP/PS1-derived mice. Plaque tissue coverage is equal between APP/PS1 and APP/PS1xTau22 mice in brain regions that accumulate fibrillar amyloid [hippocampus: t (18) = 0.32, P = 0.38; cortex: t (18) = 0.02, P = 0.16]. ( B ) Levels of non-fibrillar, soluble Aβ 1–42 species are also similar between genotypes, as measured by a human/rat-specific ELISA [ t (18) = 0.95, P = 0.18]. ( C ) AT8 immunolabelling marks extensive somatic and neuritic tau phosphorylation in CA1 regions and dentate gyrus of the hippocampus and cortex, and intensifies in amyloid-expressing animals [hippocampus: t (18) = 3.81, P = 0.0006; cortex: t (18) = 1.75, P = 0.049]. ( D and E ) Phospho-specific tau antibodies, AT100 and PHF1, label more advanced tau neurofibrillary structures in APP/PS1xTau22 mice compared with Tau22 littermates. [( D ) CA1: t (17) = 3.11, P = 0.003; cortex: t (12.6) = 2.40, P = 0.016; ( E ) CA1: t (17) = 2.49, P = 0.01; cortex: t (11.31) = 1.94, P = 0.04]. ( F ) Confocal micrographs of phospho-tau and axon-specific markers, AT8 and SMI312, respectively, show that the number of AT8 + puncta ( G ) and SMI312 + axonal swellings ( H ) are increased in the Aβ plaque core and corona when amyloid is coexpressed with pathological tau [plaque size measured: t (17) = 0.10, P = 0.46; AT8 + puncta/plaque: t (16) = 8.68, P < 0.0001; SMI312 + dystrophies/plaque: t (17) = 1.76, P = 0.048]. The plaque area measured is delineated by the dotted grey line. ns = not significant.

Article Snippet: Proteins were transferred to a nitrocellulose membrane using an iBlot transfer machine for 7 min at 25 V. Membranes were treated with LI-COR blocking buffer (LI-COR Biosciences) for 1 h at room temperature and incubated with primary antibodies against human tau (1:5000; Cat. No. A0024, DAKO) and human APP (1:5000 Cat. No. 29765, Cell Signaling), with GAPDH as a standardizing control (1:5000; Cat. No. 2118, Cell Signaling), overnight at 4°C.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Expressing

Journal: bioRxiv

Article Title: Mitochondrial accumulation and lysosomal dysfunction result in mitochondrial plaques in Alzheimer’s disease

doi: 10.1101/2025.02.19.639081

Figure Lengend Snippet: (A-B) MPs density in A and size in B at different ages of AD mice. n = 3 mice/group, including both sexes (two-tailed unpaired t-tests to compare between age-matched cortex and hippocampus in A; Ordinary one-way ANOVA, Šídák’s multiple comparisons test in B). (C) Fluorescent signals of MPs and Aβ plaques in the cortex of AD mice at different ages. White arrows: MPs without Aβ; yellow arrows: Aβ without MPs; blue arrows: scattered Aβ. (D-E) The percentage of MPs with Aβ in D and percentage of Aβ plaques with MPs in E in the cortex of AD mice at different ages. n = 3 mice/group, including both sexes (One-way ANOVA, Dunnett’s multiple comparisons test). (F) Percentages of MPs with Aβ plaques quantified based on the MPs size percentile in AD mice cortex (50 wks). n = 3 mice/group, including both sexes (one-way ANOVA, Dunnett’s multiple comparisons test). (G) Fluorescent images showing APP and mt-Keima excited at 488nm in a MP region of AD mice at 25 wks (cortex). (H) The co-localization fraction between mt-Keima and APP in MPs enriched regions of AD mice. n = 39 MPs from 3 AD mice at 25 wks, including both sexes. (I) Fluorescent images showing APP and VDAC2 in the hippocampus of AD patients. Dashed lines: MP-enriched regions; solid lines: zoom-in area. (J) Comparison of VDAC2 and APP co-localization between MP-enriched regions and non-MP regions in AD patients. n = 141 and 89 for MPs enriched regions and non-MPs regions from 4 AD individuals, respectively (Two-way ANOVA, Šídák’s multiple comparisons test). Data are mean and SEM. *P < 0.05, P** < 0.01, ***P < 0.001.

Article Snippet: The specific primary antibodies used for human brain samples include APP (Proteintech, 60342-1-Ig), VDAC2 (Proteintech, 11663-1-AP), LAMP2 (Proteintech, 66301-1-Ig), β-Amyloid (D3D2N) (Cell Signaling, 15126), MAP2 (Thermofisher Scientific, PA1-16751).

Techniques: Two Tailed Test, Comparison