Journal: bioRxiv

Article Title: Human neutralizing antibodies to cold linear epitopes and to subdomain 1 of SARS-CoV-2

doi: 10.1101/2022.11.24.515932

Figure Lengend Snippet: (A) Cartoon diagram of the SARS-CoV-2 S with highlighted coldspot regions at the fusion peptide (FP, red) and near heptad repeat 2 (HR2, blue), as well as subdomain 1 (SD1, green). The thick horizontal lines underneath indicate the location of all coldspots, and the histograms show the frequency of aa changes throughout S at different time points: 31.12.2020 (671, 006), 31.12.2021 (7,526,116) and 29.04.2022 (10,480,461 sequences from GISAID). (B) ELISA reactivity of plasma IgG antibodies to FP (top) and HR2 (middle) peptides or to SARS-CoV-2 RBD (bottom). SARS-CoV-2 (n=67) and common cold CoV (n=6) convalescent samples are compared to COVID-19 vaccinees (mRNA: BNT162b2 n=11, purple; mRNA-1273 n=5, pink; inactivated virus: Sinovac n=2, dark green; Sinopharm n=24, light green; adenovirus: ChAdOx1-S n=19, orange; Ad26.COV2.S n=4, yellow), and to non-infected controls (n=25). Area Under the Curve (AUC); average of two experiments. The percentage of samples with antibody levels 4 standard deviations above controls is shown on top. (C) Gating strategy for sorting peptide-specific or SD1-enriched B cells by flow cytometry. (D) Identification of FP-peptide specific (top) and HR2-peptide specific (bottom) B cells by flow cytometry. Numbers indicate percentage of gated double-positive cells. (E) Clonal analysis of antibody sequences derived from peptide-specific B cells identified in (D) . Pie charts show the total number of antibody sequences (center); the size of the slices is proportional to the number of clonally related sequences. White slices indicate antibody sequences that are not part of a clone.

Article Snippet: Briefly, 293T cells were transfected with pHIV NL GagPol, pCCNanoLuc2AEGFP and pSARS-CoV2-S trunc plasmids using PEI-MAX (Polysciences).

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Flow Cytometry, Derivative Assay

Journal: Cell host & microbe

Article Title: Genome-wide CRISPR screen identifies semaphorin 6A and 6B as receptors for Paeniclostridium sordellii toxin TcsL

doi: 10.1016/j.chom.2020.03.007

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Briefly, 3 × 10 7 Expi293F cells were transfected with 37.5 μg plasmid using PEIMax (1 mg/mL) (Polysciences Inc.).

Techniques: Purification, Recombinant, Modification, Protease Inhibitor, Expressing, BIA-KA, DNA Extraction, Software, Fluorescence, Microscopy, Imaging

Journal: iScience

Article Title: Activation of AMP-activated protein kinase (AMPK) through inhibiting interaction with prohibitins

doi: 10.1016/j.isci.2023.106293

Figure Lengend Snippet:

Article Snippet: HepG2 cells were seeded in a CellBIND Surface HYPERFlask (10020, Corning) and cultured for 72 h before transfection for 6 h with a mixture of 1944 μL of PEImax (PSI24765-2, Cosmo Bio, Tokyo, Japan), 423 μg of a PHB1-3×Flag expression vector, and 216 μg of a NanoLuc-AMPKα2 expression vector in DMEM containing 5.5 mM glucose.

Techniques: Produced, FLAG-tag, Control, Recombinant, Membrane, Transfection, Reverse Transcription, Silver Staining, Isolation, Cell Culture, Expressing, Plasmid Preparation, Luciferase, shRNA, Software

Journal: bioRxiv

Article Title: AAVone: A Cost-Effective, Single-Plasmid Solution for Efficient AAV Production with Reduced DNA Impurities

doi: 10.1101/2025.01.07.631712

Figure Lengend Snippet: (A) Schematic illustrations of the AAVone, triple-plasmid (standard, pTri-plasmid; and mini-pHelper, mTri-plasmid) systems, dual-plasmid (pDG-like and AAVdual) systems, and the AAVone system. (B) A comparative analysis of AAV productivity using AAVone and alternative systems in adherent HEK-293T cells. For every system compared, the total pDNA amount was maintained at 0.5 µg/well (24-well plate), and the PEImax to DNA ratio was consistently kept at 3:1. The plasmid molecular ratios for each system is displayed under each data set. (C) A comparison of vector yields with different serotypes between the AAVone and the pTri-plasmid system in suspension HEK-293T cells. The pTri-plasmid set was transfected at equal plasmid molecular ratios at a total DNA amount of 1 μg per million cells, at a cell density of 3E6 cells/mL. Crude lysate samples were quantified using qPCR with primers targeting Egfp , and data were normalized to the average values of each pTri-plasmid group. (D) Productivity optimization of the AAVone system in suspension cultured HEK-293T cells as assessed by crude lysate titration. Different cell densities, total pDNA, and transfection reagent (PEImax) were examined.

Article Snippet: The transfections were carried out using either PEImax (Polyscience, Warrington, PA) or FectoVIR (Polyplus, NY), adhering strictly to the provided guidelines.

Techniques: Plasmid Preparation, Comparison, Suspension, Transfection, Cell Culture, Titration