Transfection and Seeding of Human Tau Isoforms

Human neuroblastoma SH-SY5Y cells were maintained as described.³⁷ (link) Cells were cultured to 40–50% confluence in 6-well or 12-well plates and transfected with plasmids using X-tremeGENE^TM 9 (Roche Life Science) according to the manufacturer’s instructions. We used non-tagged human tau 3R1N, 4R1N, haemagglutinin (HA)-tagged human tau 3R1N, 4R1N, and FLAG-tagged human tau 3R1N, 4R1N in the pCDNA3.1 vector. After transfection of plasmids, cells were incubated for 6–8 h, and pathogenic tau seeds (2 μl for 6-well plate or 1–2μl for 12-well plate) were introduced using MultiFectam (Promega) according to the manufacturer’s instructions. Transfected cells were incubated for 3 days.