Plasmid Preparation:Article Title: Amyloid precursor protein is a restriction factor that protects against Zika virus infection in mammalian brains
Article Snippet: .. The pCAX FLAG APP (Addgene plasmid # 30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid # 30137), an expression plasmid for predominant form of APP in the brains, were gifts from Dennis Selkoe & Tracy Young-Pearse(39). pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein and was a gift from Dr. Asit Patnaik. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. Human pluripotent stem cells (hPSCs) differentiation into neural progenitor cells (hNPCs) and mature cortical neurons and ZIKV infection The hPSCs (H9 hESCs) were purchased from WiCell Research Institute (Cat # WA09, WiCell).
Article Title: Method to treat Alzheimer's disease
Article Snippet: The pooled human APP siRNA/shRNA/RNAi Lentivirus (siAPP; cat #: iV001195) and its control, Scrambled siRNA GFP Lentivirus (siScramble; cat #: LVP015-G) were both purchased from Applied Biological Materials Inc. .. The pCAX FLAG APP (Addgene plasmid #30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid #30137), an expression plasmid for predominant form of APP in the brains. pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. The antibodies for β-amyloid (B-4) (sc-28365), glyceraldehyde-3-phosphate dehydrogenase (0411) (sc-47724), and goat anti-mouse IgG-HRP (sc-2005) were from Santa Cruz.
Article Title: A multi‐layered network model identifies Akt1 as a common modulator of neurodegeneration
Article Snippet: HEK293 cells having 70–80% cell density were transiently transfected with pEGFP‐C1‐Aβ 1‐42 , pEGFP‐Htt‐exon1‐Q74 (Addgene #40262), pEGFP‐Ataxin1‐52Q (Addgene #32492), or pEGFP‐C1‐Ataxin3‐Q84 (Addgene #22123) DNA using Lipofectamine 2000 (Invitrogen, CA, USA) following the manufacturer's instructions. .. The pEGFP‐C1‐Aβ 1–42 plasmid was constructed from pCAX‐FLAG‐APP (Addgene #30154). .. Before drug treatment, HEK293 cells were washed with treating medium (Minimum essential medium (MEM, Gibco, MD, USA) with 1% FBS) and then treated with the indicated concentration of SC79 (0.1–100 μM) (SML0749, Sigma‐Aldrich) for 24 h. Cell death was measured using Cell Counting Kit‐8 (CK04, Dojindo, MD, USA), which was performed according to the manufacturer's instructions.
Article Title: Methods of treating Alzheimers disease
Article Snippet: Both secondary antibodies were used with 1:1000 dilutions and the dilution buffer for primary and secondary antibodies was 0.1% Tween 20 (Sigma) and 0.4% bovine serum albumin (BSA; A7906; Sigma) in 1×PBS. .. The pCAX FLAG APP (Addgene plasmid #30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid #30137), an expression plasmid for predominant form of APP in the brains. pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. All samples were examined a Nikon-Ti2 fluorescence microscope and images were obtained using a Nikon A1r-Ti2 confocal system at the Microscopy Core Facility at the UNL.
Expressing:Article Title: Amyloid precursor protein is a restriction factor that protects against Zika virus infection in mammalian brains
Article Snippet: .. The pCAX FLAG APP (Addgene plasmid # 30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid # 30137), an expression plasmid for predominant form of APP in the brains, were gifts from Dennis Selkoe & Tracy Young-Pearse(39). pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein and was a gift from Dr. Asit Patnaik. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. Human pluripotent stem cells (hPSCs) differentiation into neural progenitor cells (hNPCs) and mature cortical neurons and ZIKV infection The hPSCs (H9 hESCs) were purchased from WiCell Research Institute (Cat # WA09, WiCell).
Article Title: Method to treat Alzheimer's disease
Article Snippet: The pooled human APP siRNA/shRNA/RNAi Lentivirus (siAPP; cat #: iV001195) and its control, Scrambled siRNA GFP Lentivirus (siScramble; cat #: LVP015-G) were both purchased from Applied Biological Materials Inc. .. The pCAX FLAG APP (Addgene plasmid #30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid #30137), an expression plasmid for predominant form of APP in the brains. pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. The antibodies for β-amyloid (B-4) (sc-28365), glyceraldehyde-3-phosphate dehydrogenase (0411) (sc-47724), and goat anti-mouse IgG-HRP (sc-2005) were from Santa Cruz.
Article Title: Methods of treating Alzheimers disease
Article Snippet: Both secondary antibodies were used with 1:1000 dilutions and the dilution buffer for primary and secondary antibodies was 0.1% Tween 20 (Sigma) and 0.4% bovine serum albumin (BSA; A7906; Sigma) in 1×PBS. .. The pCAX FLAG APP (Addgene plasmid #30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid #30137), an expression plasmid for predominant form of APP in the brains. pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. All samples were examined a Nikon-Ti2 fluorescence microscope and images were obtained using a Nikon A1r-Ti2 confocal system at the Microscopy Core Facility at the UNL.
Cloning:Article Title: Amyloid precursor protein is a restriction factor that protects against Zika virus infection in mammalian brains
Article Snippet: .. The pCAX FLAG APP (Addgene plasmid # 30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid # 30137), an expression plasmid for predominant form of APP in the brains, were gifts from Dennis Selkoe & Tracy Young-Pearse(39). pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein and was a gift from Dr. Asit Patnaik. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. Human pluripotent stem cells (hPSCs) differentiation into neural progenitor cells (hNPCs) and mature cortical neurons and ZIKV infection The hPSCs (H9 hESCs) were purchased from WiCell Research Institute (Cat # WA09, WiCell).
Article Title: Method to treat Alzheimer's disease
Article Snippet: The pooled human APP siRNA/shRNA/RNAi Lentivirus (siAPP; cat #: iV001195) and its control, Scrambled siRNA GFP Lentivirus (siScramble; cat #: LVP015-G) were both purchased from Applied Biological Materials Inc. .. The pCAX FLAG APP (Addgene plasmid #30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid #30137), an expression plasmid for predominant form of APP in the brains. pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. The antibodies for β-amyloid (B-4) (sc-28365), glyceraldehyde-3-phosphate dehydrogenase (0411) (sc-47724), and goat anti-mouse IgG-HRP (sc-2005) were from Santa Cruz.
Article Title: Methods of treating Alzheimers disease
Article Snippet: Both secondary antibodies were used with 1:1000 dilutions and the dilution buffer for primary and secondary antibodies was 0.1% Tween 20 (Sigma) and 0.4% bovine serum albumin (BSA; A7906; Sigma) in 1×PBS. .. The pCAX FLAG APP (Addgene plasmid #30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid #30137), an expression plasmid for predominant form of APP in the brains. pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. All samples were examined a Nikon-Ti2 fluorescence microscope and images were obtained using a Nikon A1r-Ti2 confocal system at the Microscopy Core Facility at the UNL.
Transfection:Article Title: Amyloid precursor protein is a restriction factor that protects against Zika virus infection in mammalian brains
Article Snippet: .. The pCAX FLAG APP (Addgene plasmid # 30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid # 30137), an expression plasmid for predominant form of APP in the brains, were gifts from Dennis Selkoe & Tracy Young-Pearse(39). pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein and was a gift from Dr. Asit Patnaik. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. Human pluripotent stem cells (hPSCs) differentiation into neural progenitor cells (hNPCs) and mature cortical neurons and ZIKV infection The hPSCs (H9 hESCs) were purchased from WiCell Research Institute (Cat # WA09, WiCell).
Article Title: Method to treat Alzheimer's disease
Article Snippet: The pooled human APP siRNA/shRNA/RNAi Lentivirus (siAPP; cat #: iV001195) and its control, Scrambled siRNA GFP Lentivirus (siScramble; cat #: LVP015-G) were both purchased from Applied Biological Materials Inc. .. The pCAX FLAG APP (Addgene plasmid #30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid #30137), an expression plasmid for predominant form of APP in the brains. pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. The antibodies for β-amyloid (B-4) (sc-28365), glyceraldehyde-3-phosphate dehydrogenase (0411) (sc-47724), and goat anti-mouse IgG-HRP (sc-2005) were from Santa Cruz.
Article Title: Methods of treating Alzheimers disease
Article Snippet: Both secondary antibodies were used with 1:1000 dilutions and the dilution buffer for primary and secondary antibodies was 0.1% Tween 20 (Sigma) and 0.4% bovine serum albumin (BSA; A7906; Sigma) in 1×PBS. .. The pCAX FLAG APP (Addgene plasmid #30154) is a FLAG-tagged APP expression plasmid APP and pCAX APP 695 (Addgene plasmid #30137), an expression plasmid for predominant form of APP in the brains. pCI-neo-ZIKV-prME is expression plasmid for PrM-E protein. pcDNA3 (Invitrogen) is a cloning vector used as controls for transfections. .. All samples were examined a Nikon-Ti2 fluorescence microscope and images were obtained using a Nikon A1r-Ti2 confocal system at the Microscopy Core Facility at the UNL.
Construct:Article Title: A multi‐layered network model identifies Akt1 as a common modulator of neurodegeneration
Article Snippet: HEK293 cells having 70–80% cell density were transiently transfected with pEGFP‐C1‐Aβ 1‐42 , pEGFP‐Htt‐exon1‐Q74 (Addgene #40262), pEGFP‐Ataxin1‐52Q (Addgene #32492), or pEGFP‐C1‐Ataxin3‐Q84 (Addgene #22123) DNA using Lipofectamine 2000 (Invitrogen, CA, USA) following the manufacturer's instructions. .. The pEGFP‐C1‐Aβ 1–42 plasmid was constructed from pCAX‐FLAG‐APP (Addgene #30154). .. Before drug treatment, HEK293 cells were washed with treating medium (Minimum essential medium (MEM, Gibco, MD, USA) with 1% FBS) and then treated with the indicated concentration of SC79 (0.1–100 μM) (SML0749, Sigma‐Aldrich) for 24 h. Cell death was measured using Cell Counting Kit‐8 (CK04, Dojindo, MD, USA), which was performed according to the manufacturer's instructions.
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