Primary embryonic mouse neuron cultures were prepared from freshly dissected embryonic cortices, as described previously. Briefly, pregnant female CD-1 mice (Charles River) were euthanized at embryonic days 14 to 16, embryonic cortices were dissected and homogenized using a papain dissociation kit (Worthington Biochemical), and neurons were plated on culture dishes that were pretreated with poly-l-lysine. For tau propagation studies, neurons were transduced by the direct addition of AAV enhanced GFP (eGFP)–2a-WTtau or eGFP-2a-P301Ltau particles to the culture medium at 7 days in vitro (DIV), and neurons were fixed for immunostaining at 14 DIV or detached with StemPro Accutase Cell Dissociation Reagent (Thermo Fisher Scientific) from the culture dish for flow cytometry at 14 or 21 DIV.

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