Journal: bioRxiv

Article Title: Cell specificity of adeno-associated virus (AAV) serotypes in human cortical organoids

doi: 10.1101/2023.04.13.536491

Figure Lengend Snippet: ( a & b ) Example of immunohistological sample of Iso clone, 42 days post transduction with AAV6 and labeled with either (a) GFP, MAP2, and DAPI or (b) GFP, GFAP, and DAPI. Dotted square indicates enlarged area. White arrows indicate co-staining of GFP with either (b) MAP2 or (b) GFAP. All hCOs were harvested on day 120, 10 days post AAV transduction. ( c ) Flow cytometry quantification of % GFP labeled cells for each AAV serotype in each hCO clone. ( d ) Flow cytometry quantification of % GFP and MAP2 co-labeled cells for each AAV serotype in each hCO clone. ( e ) Flow cytometry quantification % GFP and GFAP co-labeled cells for each AAV serotype in each hCO clone. All hCOs were harvested on day 120, 10 days post AAV transduction. ( n = 4 hCOs per condition, statistical significance between AAV6 and AAV2 or AAV9 serotypes determined by one-way ANOVA with Tukey post-hoc tests, *** p < 0.001, ** p < 0.01. Values for comparisons between other serotypes are in Supplementary Tables 1-9).

Article Snippet: For staining, slides were rinsed with PBS, then incubated in 0.25% Triton X-100 with 4% Donkey serum (EMD Millipore, S30) for 1 hour, followed by incubation in primary antibodies overnight: chicken GFP (1:3000, Aves Labs, GFP-1010) and guinea pig MAP2 (1:1000, Millipore-Sigma, MAB3418).

Techniques: Transduction, Labeling, Staining, Flow Cytometry