tau rd p301s biosensor cell culture  (ATCC)

Journal: Alzheimer's & Dementia

Article Title: ApoE3 Christchurch and tau interaction as a protective mechanism against Alzheimer's disease

doi: 10.1002/alz.70396

Figure Lengend Snippet: ApoE3 WT and ApoE3Ch differentially interacts with tau. (A, B) Number of MAPT fragments and sum intensity obtained via mass spectrometry analysis (* p ≤ 0.05, ** p ≤ 0.01, Two‐way ANOVA, Fisher's LSD post‐hoc test, n = 3). (C) Co‐immunoprecipitation of human tau recombinant protein using His‐tag ApoE3 WT and ApoE3Ch proteins and detected via WB using anti‐tau antibody. (D) Representative Western blot of ApoE3 WT and ApoE3Ch after incubation with monomeric recombinant human tau or tau PFFs, arrows point the different tau aggregate bands. (E) Levels of FRET signal in tau RD P301S FRET Biosensor cells stimulated with tau PFF and treated with APOE3 variants (* p ≤ 0.05, ** p ≤ 0.01, one‐way ANOVA, Fisher's LSD post‐hoc test, n = 6). (F) Scale bar: 100 µm. Representative maximum intensity projections of the live imaging acquisition of the tau aggregates in the tau RD P301S FRET Biosensor cells stimulated with tau PFFs and treated with the ApoE3 variants. (G) WB of tau aggregates in tau RD P301S FRET Biosensor cells treated with ApoE3 variants and ratio quantification between the total tau aggregates at ∼ 150 kDa of molecular weight and GAPDH, quantified bands are indicated by an arrowhead. (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, one‐way ANOVA, Fisher's LSD post‐hoc test, n = 6). ANOVA, analysis of variance; ApoE3, apolipoprotein E3; ApoE3Ch, apolipoprotein E3 Christchurch variant; GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase; LSD, least significant difference; PFF, preformed protofibrils; WB, Western blotting; WT, wild‐type.

Article Snippet: Tau RD P301S FRET Biosensor cells (ATCC, Manassas, VA, cat. CRL‐3275) were cultured in DMEM with glutaMAX (Gibco, Waltham, MA, cat. 10566016) and 10% fetal bovine serum (FBS) until confluency.

Techniques: Mass Spectrometry, Immunoprecipitation, Recombinant, Western Blot, Incubation, Imaging, Molecular Weight, Variant Assay

Journal: Alzheimer's & Dementia

Article Title: ApoE3 Christchurch and tau interaction as a protective mechanism against Alzheimer's disease

doi: 10.1002/alz.70396

Figure Lengend Snippet: Effect of Christchurch variant on ApoE‐induced tau pathology in the mouse retina. (A) Representative immunofluorescence of subretinally injected MAPT P301S tau transgenic mice with recombinant ApoE3 and ApoE3Ch variants, as compared to vehicle, showing reduced levels of AT8‐positive cells in the presence of the Ch variant. (B) Relative AT8+ tau levels in dissected retina isolated from mice treated with either vehicle, ApoE3, or ApoE3Ch, showing that in the presence of ApoE3Ch, AT8+ cells are significantly reduced (** p < 0.005, One‐way ANOVA, n = 3‐4). (C) immunofluorescence staining of intravitreal injected 9 months old MAPT P301S tau mice with recombinant ApoE3 and ApoE3 Ch proteins, showing the reduced levels of ptau S396‐positive cells in the presence of the Ch variant. (D) Intravitreal injection of ApoE3 Ch in the retina significantly decreased phosphorylation of tau (* = p < 0.05, one‐way ANOVA, n = 2–5). Vehicle treated wild‐type mice were used as control for MAPT P301S mice (PS19). For both panels A and C, nuclei are stained with DAPI in blue, vascular formation is stained with Isolectin (IB4) in green and tau phosphorylation are stained with AT8 anti tau (A) or ptau S396 antibody (C) in red. Scale bar = 50 µm. ANOVA, analysis of variance; ApoE3, apolipoprotein E3; ApoE3Ch, apolipoprotein E3 Christchurch variant; DAPI, 4′,6‐diamidino‐2‐phenylindole.

Article Snippet: Tau RD P301S FRET Biosensor cells (ATCC, Manassas, VA, cat. CRL‐3275) were cultured in DMEM with glutaMAX (Gibco, Waltham, MA, cat. 10566016) and 10% fetal bovine serum (FBS) until confluency.

Techniques: Variant Assay, Immunofluorescence, Injection, Transgenic Assay, Recombinant, Isolation, Staining, Phospho-proteomics, Control