Mammalian tissue culture

SHSY5Y cells were obtained from the Duke University Cell Culture Facility and were maintained at 37 °C with 5% CO₂ in DMEM:F12 (1:1; Gibco) supplemented with 10% FBS (Sigma Aldrich) and 1X penicillin-streptomycin (Gibco). Cells were passaged at 80% confluency with a sub-cultivation ratio of 1:20 unless seeding for transfection experiments where a higher density was required. Transfection of SHSY5Y was performed with GeneX Plus (ATCC). Transfected plasmids are described in Supplemental Table 1.