Journal: Cell reports

Article Title: Evidence for a clathrin-independent endocytic pathway for APP internalization in the neuronal somatodendritic compartment

doi: 10.1016/j.celrep.2023.112774

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal Anti-Bace1 , ThermoFisher , Cat#PA1–757; RRID:AB_325863.

Techniques: Virus, Recombinant, In Situ, Enzyme-linked Immunosorbent Assay, BIA-KA, Plasmid Preparation, Software

Journal: International Journal of Nanomedicine

Article Title: A method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency

doi: 10.2147/IJN.S78935

Figure Lengend Snippet: Nanocomplexes achieve silencing ex vivo, but not in the brain following intravenous administration. Notes: Mice brain explants were transfected ex vivo with peptide/siRNA complexes (PR: RVG-9R/BACE1siRNA; 4:1 weight ratio), anionic PEGylated PRL nanocomplexes with BACE1 siRNA, L2K, or complexes with irrelevant control siRNA (all at 100 nM), and then 48 hours post-transfection, the tissues were processed for analysis ( A ) by qRT-PCR and ( B ) by Western blot analysis of the BACE1 protein. Protein silencing was calculated with densitometric analysis using tubulin as loading control. ( C ) Anionic PRL nanocomplexes containing RVG-9R and BACE1 siRNA were concentrated using 300 g/L dextran over 3.5 hours, and this concentrated nanoparticle formulation was used ( D ) in intravenous injections. Mice were injected with 100 μL of anionic PRL nanoparticles containing 16 μg or 50 μg BACE1 siRNA or IRR siRNA, and 48 hours later, brains were processed for qRT-PCR analysis. The values are the means of three animals ± standard deviation. Asterisks indicate comparisons of specific formulations with statistical significance (* P <0.05; ** P <0.01). Abbreviations: mRNA, messenger RNA; IRR, irrelevant control; siRNA, small interfering RNA; L2K, Lipofectamine ® 2000; PRL, peptide Y or RVG-9R, siRNA, liposome L AP 2; RVG, rabies virus glycoprotein targeting peptide; PR, RVG-9R/BACE1siRNA; PEG, polyethylene glycol; qRT-PCR, quantitative reverse transcription polymerase chain reaction.

Article Snippet: The primary antibodies used in this study were rabbit anti-BACE1 polyclonal antibody (EE-17, 1:1,000; Sigma-Aldrich Co.) and mouse anti-β-tubulin monoclonal antibody (1:5,000; Sigma-Aldrich Co.), and the secondary antibodies were a horseradish peroxidase-conjugated antimouse or antirabbit immunoglobulin G (Stratech Scientific Ltd, Suffolk, UK) (1:50,000).

Techniques: Ex Vivo, Transfection, Quantitative RT-PCR, Western Blot, Injection, Standard Deviation, Small Interfering RNA, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Nanomedicine

Article Title: A method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency

doi: 10.2147/IJN.S78935

Figure Lengend Snippet: In vivo silencing of BACE1 following CED administration of PEGylated concentrated nanoparticles or control saline into rat striatum. Notes: Concentrated anionic PEGylated PRL nanoparticles containing RVG-9R peptide and BACE1 siRNA or irrelevant control siRNA were administered by CED in the striatum of rats, and 48 hours postadministration, tissues were removed for qRT-PCR analysis of siRNA-induced silencing of the BACE1 gene. Values are the means of five animals ± standard deviation (n=3 for the untreated control animals and n=4 for the saline control animals) with one-way analysis of variance and Bonferroni’s post hoc analysis performed to calculate significant differences (* P <0.05; ** P <0.01; *** P <0.001). Abbreviations: mRNA, messenger RNA; siRNA, small interfering RNA; CED, convection-enhanced delivery; PEG, polyethylene glycol; PRL, peptide Y or RVG-9R, siRNA, liposome L AP 2; qRT-PCR, quantitative reverse transcription polymerase chain reaction; n, number.

Article Snippet: The primary antibodies used in this study were rabbit anti-BACE1 polyclonal antibody (EE-17, 1:1,000; Sigma-Aldrich Co.) and mouse anti-β-tubulin monoclonal antibody (1:5,000; Sigma-Aldrich Co.), and the secondary antibodies were a horseradish peroxidase-conjugated antimouse or antirabbit immunoglobulin G (Stratech Scientific Ltd, Suffolk, UK) (1:50,000).

Techniques: In Vivo, Quantitative RT-PCR, Standard Deviation, Small Interfering RNA, Convection, Reverse Transcription Polymerase Chain Reaction