Fig. 3

Effects of mitochondrial complex I inhibitors MPP⁺ and rotenone on Ser129-phosphorylation of α-syn. Cell lysates (10 μg/lane) were loaded on SDS-PAGE and analyzed by western botting with EP1536Y, Syn-1, or AC-15 antibody. a Effect of incubation time of MPP⁺ and rotenone on Ser129-phosphorylation. Wt-aS/SH #4 cells were incubated in media containing 1 mM MPP⁺ or 5 μM rotenone for the indicated time points until 16 h. Vehicle controls were treated with DMSO at the same final concentration as each reagent. b Effect of concentrations of MPP⁺ or rotenone on Ser129-phosphorylation. Cells were incubated in media containing the indicated amounts of MPP⁺ or rotenone for 12 h. Graphs show relative band intensities of Ser129-phosphorylated α-syn and total α-syn. They were normalized to those of β-actin. Data represent means ± SD and P values were estimated by one-way ANOVA with Bonferroni correction or Welch-ANOVA with Games-Howell post hoc test for unequal-variances (*, P < 0.05; **, P < 0.01)