2.5. Establishment of SH-SY5Y-A53T and Cell Culture

We purchased the SH-SY5Y cell line from the American type culture collection (ATCC, CRL-2266, USA). The cell line was transfected by lentivirus particles stably overexpressing SNCA (A53T) after puromycin-based screening. In the α-synuclein A53T mutant gene, the amino acid at position 53 was converted from alanine (A) to threonine (T) as a result of a missense mutation at position 209 from G to A [34]. Cells were cultured in Dulbecco's modified Eagle's medium (DMEM, GIBCO) containing 10% fetal bovine serum (FBS, GIBCO, Australia).