Fig. 3

Transduction of BSC by secreted rAAV from select capsids. Organotypic BSCs were prepared and transduced at 0 DIV with rAAV (1 × 10¹⁰ vg/mL of BSC culture media). BSCs were maintained in culture until 14 DIV, fixed, and fluorescence of EGFP throughout the BSC was imaged. Representative images of BSC transduction by rAAV2-EGFP vectors pseudotyped with wild-type capsids (a) or mutant capsids (b) Bar, 100 μm. Images were taken with the same exposure time to directly compare transduction efficiency. c BSCs were transduced with rAAV2/8-WT-htau or rAAV2/8-P301L-S320F-htau under the hCBA promoter (1 × 10¹⁰ vg/mL of BSC culture media) at 0 DIV and maintained in culture until 28 DIV. Lysates were sequentially extracted and immunoblotted for total human tau (CP27), tau phosphorylated at Ser396/404 (PHF-1), and GAPDH as a loading control. Representative western blots of tau in the low-speed supernatant and sarkosyl insoluble fraction are shown. e Transduced slice cultures were fixed, stained with 0.0125% Thioflavin S, and imaged to identify any β-sheet structures in these sections Bar, 25 μm