Figure 2.

Effects of PLD1 inhibition on the accumulation of α-synuclein aggregates. (a) PLD1 inhibition induced accumulation of α-synuclein. Differentiated SH-SY5Y cells overexpressing human α-synuclein were treated with 20 μM of the PLD1 inhibitor for 2 days. The levels of α-synuclein were normalized with β-actin. HM, high molecular species; M, Monomeric size. N=3, *P<0.05. (b) α-Synuclein accumulation in APs. Differentiated SH-SY5Y cells were co-transfected with human α-synuclein and tfLC3 and incubated with the PLD1 inhibitor as above. Cells were permeabilized with 40 μM of digitonin before immunofluorescence staining. A total of 300 cells were analyzed from three independent experiments (100 cells per experiment). Scale bar: 20 μm, *P<0.05. (c) Immunogold EM of α-synuclein in APs in cells treated with PLD1 inhibitor. N=3 (15 cells per experiment). *P<0.05. (d) Cytotoxicity analysis. Control cells and cells overexpressing human α-synuclein were cultured for 1 day and treated with either DMSO or PLD1 inhibitor for 2 days. N=3, *P<0.05, ^#P<0.05