Fig. 3. DAPK1 promotes α-synuclein aggregation in SH-SY5Y cells. (A) After SH-SY5Y cells were transfected for 24 h with plasmids encoding Myc-α-synuclein alone or with increasing amounts of FLAG-DAPK1-WT or FLAG-DAPK1-K42A, filter trap assay of cell lysates was performed. (B) SH-SY5Y cells were transfected for 24 h with plasmids encoding Myc-α-synuclein or Myc-α-synuclein-S129A, or FLAG-DAPK1 alone or in combination. Cell lysates were prepared, and filter trap assay was performed. (C) Representative confocal images of the immunostaining of GFP-α-synuclein (green) and FLAG-DAPK1 (red) are shown. The number of cells showing visible α-synuclein aggregates versus the number showing no-apparent granular inclusions was calculated and the percent ratio is shown as a graph. (D) Representative confocal images of the immunostaining of pS129-α-synuclein (green) and FLAG-DAPK1 (red) are shown. The number of cells showing visible α-synuclein aggregates vs the number of cells showing no-apparent granular inclusions was calculated, and their percent ratios are shown as a graph. All graphed data represent the mean±SEM of five independent experiments (**p≤0.01; a-d).
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