Journal: Cell reports

Article Title: Caspase Activation and Caspase-Mediated Cleavage of APP Is Associated with Amyloid β-Protein-Induced Synapse Loss in Alzheimer’s Disease

doi: 10.1016/j.celrep.2020.107839

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-GFP Aves Lab #GFP-1020; RRID:AB_10000240 CT15 UCSD APP680-695 6E10 Signet Laboratories S G-39320; RRID:AB_662798 82E1 IBL international JP10323; RRID:AB_1630806 β-Tubulin Calbiochem CP07; RRID:AB_565242 p-GluR1 Thermo Fisher 36-8300; RRID:AB_2533280 NMDA R-1 Novus NBP1-48559; RRID:AB_10011203 activated caspase-3 Ab Cell Signaling 9664; RRID:AB_2070042 β-actin Sigma-Aldrich A5316; RRID:AB_476743 HRP-conjugated 6E10 Covance SIG-39320; RRID:AB_662798 Bacterial and Virus Strains Sindbis Virus-GFP UCSD N/A Sindbis Virus-Tomato UCSD N/A Sindbis Virus-C99ΔC UCSD N/A Sindbis Virus-C99ΔC_Tomato UCSD N/A mMessage mMachine SP6 Ambion, Life Technologies AM1340 Chemicals, Peptides, and Recombinant Proteins DEVD-ultrabright green dye (FLICA) Vergent Bioscience 13101 Hanks’ Balanced Salt Solution GIBCO 24020-117 heat inactivated horse serum GIBCO 16050-122 MEM-HEPES GIBCO 12360-038 PG-SM GIBCO 15140-148 GlutaMAX Sigma-Aldrich G2150 DAPT EMD Biosciences 565770 zVAD R&D Systems FMK001 zDEVD R&D Systems FMK004 zYVAD R&D Systems FMK005 zVEID R&D Systems FMK006 zIETD R&D Systems FMK007 active caspase 3 protein Salvesen’s lab, SBP N/A Vector shield vector laboratories H-1000 Harleco’s Krystalon mounting medium Millipore 6496964969 protease inhibitor Sigma-Aldrich S8820 Critical Commercial Assays TranslT LT-1 Transfection reagent Mirus MIR2304 Experimental Models: Organisms/Strains B6.129S4-Gt(ROSA)26Sortm2(FLP*)Sor/J Jackson Laboratory #012930 GFP-M Jackson Laboratory #007788 APP D664A KI UCSD N/A Hippocampal organotypic slice 5–7 day old mouse hippocampi culture, UCSD N/A Oligonucleotides GFP forward primer (AAG TTC ATC TGC ACC ACC G) N/A GFP reverse primer (TCC TTG AAG ATG GTG CG) N/A APP D664A AMBE4 (ATG AAC ACC GAT GG G TAG TGA AGC) N/A APP D664A WT16 (TGC TTT CTA GGT CGA C) N/A APP D664AKI16 (TGC TTT CTA GGT CGC T) N/A Software and Algorithms pClamp 10 Molecular Devices N/A NeuronStudio BISE N/A Imaris Bitplane N/A ImageJ NIH https://imagej.nih.gov/ij/ Prism GraphPad N/A Other FBS Omega FB-02 DMEM Corning Cellgro MT10013CV Cell Culture Inserts Millicell® PICM0RG50 Centriprep filters Amicon, Beverly 4303 glass micropipette Warner Instruments N/A Picospritzer II General Valve Corporation N/A SD9 Stimulator Grass Technologies N/A Open in a separate window KEY RESOURCES TABLE Aβ-dependent caspase activation cleaves the APP C terminus, leading to synaptic injury Inhibition of APP C-terminal cleavage ameliorates Aβ-dependent synaptic injury APP D664A mutation prevents APP C terminus cleavage and impairs caspase activation Synaptic injury by Aβ is rescued in APP D664A knock-in mice

Techniques: Virus, Recombinant, Plasmid Preparation, Protease Inhibitor, Transfection, Software, Cell Culture