vsv-g envelope protein plasmid pmd2g  (Addgene inc)

Journal: bioRxiv

Article Title: NK cell immunotherapy administered at the time of HIV recrudescence is associated with viral control

doi: 10.1101/2025.11.19.688897

Figure Lengend Snippet: A) Schematic figure showing the overview of NK cell production and the overview of the mouse study timeline. Red circles on the timeline indicate blood collection, black dots indicate scheduled necropsies. Schematic of transgene design: MND promoter, CAR (CD4-MBL-CD8TM-4-1BB-CD3z), CXCR5, IL-15, the BgH PolyA tail, with separation mediated by P2A and T2A sites, and inverted terminal repeats (ITRs). Schematic showing expression of all transgene components as well as base editor-mediated knock out of PD-1 on the NK cell surface. Figure made using Biorender.com. B) Expression of CAR (MBL) and CXCR5 molecules was confirmed in the cell product using flow cytometry. Cells were pre-gated sequentially on lymphocytes, singlets, live cells, CD56+, and CD3-. C) PD-1 knockout was detected at the RNA level by RT-PCR, CAR NK (blue) relative to control NK cells (black). D) IL-15 expression in CAR NK (blue) and control NK (black) cells via IL-15 ELISA of cell culture supernatant. E) CAR (blue) and control (black) cell proliferation over one week post-thaw. F) The specific lysis of HIV Envelope P815s by CAR (blue) or control (black) NK cells via DELFIA cytotoxicity assay. 5:1, 10:1, and 20:1 E:T ratios were tested. All assays were performed in duplicate or triplicate. Data are represented as mean + standard deviation (error bars).

Article Snippet: Inducible truncated HIV-Env P815s-The P815 truncated HIV Env cell line was produced by retronectin-mediated transduction of P815 cells (a gift from Dr. Geoffery Hart) using a lentivirus produced from a tet-inducible truncated HIV Envelope plasmid (kindly provided by Dr. Alon Herschhorn; Tet-One Inducible Expression System, Clontech) and psPax2 and VSV-G plasmids.

Techniques: Expressing, Knock-Out, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Control, Enzyme-linked Immunosorbent Assay, Cell Culture, Lysis, Cytotoxicity Assay, Standard Deviation

Journal: bioRxiv

Article Title: Concerted remodelling of the postsynaptic spine and RNA granule by cLTP

doi: 10.1101/2025.07.16.665171

Figure Lengend Snippet: a , Experimental design of biotinylation assays in cortical neurons under basal, cLTP and post-cLTP conditions. b , A representative αFLAG immunofluorescence image of cortical neurons expressing FLAG-APEX2 fused to a nuclear export signal sequence from a lentiviral vector. c , Proteomic analysis of streptavidin pulldown and input samples from cortical neurons subject to biotin labelling under basal conditions. d , Pulldown to input ratios as a function of the predicted number of surface tyrosines per protein. e , Volcano plot with enrichment scores for the indicated selection of protein sets (bubble size indicates protein number in each protein set). f , Tyrosine surface exposure in ribosomal proteins with low and high accessibility scores as determined by their streptavidin pulldown / input ratios. g , Accessibility scores corresponding to ribosomal proteins as a function of the number of surface tyrosines hidden in the 80S ribosome under basal (blue) and cLTP (orange) conditions. h , Proteomic analysis of cortical neurons under basal and cLTP conditions. Ribosomal proteins are indicated (red dots).

Article Snippet: HEK293T cells were transfected with pFUW lentiviral vectors, envelope plasmid pVSV-G (Addgene, 8454) and packaging plasmid pCMV-dR8.2 dvpr (Addgene 8455).

Techniques: Immunofluorescence, Expressing, Sequencing, Plasmid Preparation, Selection