Journal: eLife

Article Title: Molecular and anatomical organization of the dorsal raphe nucleus

doi: 10.7554/eLife.46464

Figure Lengend Snippet:

Article Snippet: Genetic reagent (Adeno-associated virus) , AAV2/1-CAG-FLEX-tdTomato , Penn Vector Core , RRID: Addgene_51503 , 10 12 GC/ml.

Techniques: Virus, Plasmid Preparation, Generated, Recombinant, RNAscope, Multiplex Assay, Detection Assay, Software

Journal: Nature Communications

Article Title: Modulating nuclear stiffness and envelope barrier facilitates AAV nuclear entry and reduces immunogenicity

doi: 10.1038/s41467-026-71194-5

Figure Lengend Snippet: a Immunization protocol design diagram and schematic diagram of peripheral immune organ detection indicators. b Serum anti-AAV capsid IgG, IgG2a, IgG1 titers ( n = 4). Analysis of AAV capsid-specific cellular immune response in the spleen. Flow cytometric analysis of Th1 cells ( c n = 4), CD8 + cytotoxic T cells ( d , e n = 4), AAV6-capsid-specific polyfunctional CD4 + ( f n = 4) and CD8 + memory T cells ( g , h n = 4). IFN-γ spot-forming units (SFU) of splenocytes stimulated with MHC class I AAV6-capsid-immunodominant peptide ( i n = 4). Radar map of cytokines in the lymphocytes culture medium ( j n = 4). k AAV redosing experimental scheme diagram. l Serum anti-AAV capsid IgG titers at times indicated ( n = 4). Anti-AAV IgG antibody titers following the repeat administration are presented as the ratio to the titer measured prior to the repeat dose (Day 38). m , n AAV capsid-mediated in vitro cytotoxicity assay. Female C57BL/6J mice were intramuscularly injected with AAV2-GFP. After 21 days of treatment, splenic lymphocytes from AAV2-GFP immunized mice were co-cultured with AAV2-Luc transduced Hepa1-6 cells (with or without PF127) at a ratio of 50:1 for 5 h. The levels of LDH in the cell culture supernatant were measured to calculate specific lysis (%) ( n = 7). o , p ELISA of ubiquitinated AAV2 capsid proteins in HeLa cells after transduction with AAV2-GFP or PF127 + AAV2-GFP ( n = 4). Values represent mean \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm \,$$\end{document} ± SEM: * p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$ < \,$$\end{document} < 0.05; ** p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$ < \,$$\end{document} < 0.01; *** p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$ < $$\end{document} < 0.001; and **** p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$ < $$\end{document} < 0.0001. P Values were determined by one-way ANOVA test with Tukey’s multiple comparisons test ( b ). P Values were determined by one-way ANOVA test with Dunnett’s multiple comparisons test ( c – i ). P Values were determined by Two-tailed unpaired t -test ( n , p ). Created with BioRender.com ( a , k , m , o ). Source data are provided as a file.

Article Snippet: After preclearing, 2 μg of capsid antibody against intact AAV2 particles (anti-AAV2 mouse recombinant, A20R, PROGEN #690298S) were added and incubated at 4 °C for 24 h, followed by precipitation with protein G-agarose beads.

Techniques: In Vitro, Cytotoxicity Assay, Injection, Cell Culture, Lysis, Enzyme-linked Immunosorbent Assay, Transduction, Two Tailed Test