Figure 7.

Activity of α-synuclein aggregation inhibitors assessed using ASYN-CONA: (A) mean bead ring intensity quantified from the Cy5 fluorescence detection channel and (B) ratio of the Cy5 fluorescence intensity to the TMR fluorescence intensity. First, Ni-NTA agarose beads were incubated with α-synuclein-A91C-HTM (100 nM), shaken for 20 min at 22 °C, and washed. α-Synuclein-A91C-Cy5 (500 nM) and the inhibitor (50 μM) in EtOH (20%) were added to the solution and the beads incubated for 5 h. One aliquot was taken after 5 h for imaging and fluorescence intensity quantification. Three controls with, respectively, no on-bead α-synuclein-A91C-HTM, no EtOH, or no inhibitor present in the reaction sample were also included in the assay. All experiments were performed in duplicate, and the full experiment was repeated twice. The bars in graphs represent the average weighted by the number of beads of the four repetitions.