(A) Schematic of the primary neuron seeding assay, using P301S tau transgenic mice at P2. Cultures were infected with AAV PHP.eB carrying venus-P2A-tau-RING (VPTR) or venus only at day in vitro 2 (DIV2), and P301S tau aggregates were added to the media at DIV7. Cultures were evaluated at DIV14 for the number of AT8-positive tau aggregates.
(B) Quantification of primary neurons treated ±100 nM tau aggregates, ±venus, or venus-P2A-tau-RING (VPTR) AAV at DIV14. N = 3.
(C) Representative immunofluorescence images of primary neuron cultures at DIV14, treated with venus or VPTR AAV, +100 nM P301S tau aggregates. Scale bar, 100 μm. Statistical significance for (B) determined by two-way ANOVA and Sidak’s multiple comparisons test. ****p < 0.0001. ns, non-significant.
See also Figure S3.