Fig. 5.

TLR2 activation and treatment with α-syn PFF inhibit autophagy in PD midbrain cells. PD patient iPSCs were differentiated into a midbrain model and cultured for 7 and 14 days with or without 1 μg/ml Pam3CSK4 and/or 1 μg/ml PFFs. a Cells were fixed at days indicated and stained for autophagy marker P62 (green) and MAP2 (red). Confocal images were taken at 60 × magnification with 8 images captured per condition and used for the analysis of P62 signal intensity. Scale bar, 10 μm. b Graph shows the fold change in P62 intensity/cell as normalised to the control as mean ± SEM (n = 6–9). c Cells were stained with Lysotracker (red) for 1 h at 37 °C prior to fixation at day 2 and 7 post treatment, and then co-stained for MAP2 (green). Confocal images were taken at 60 × magnification with 8 images captured per condition and used for the analysis. Scale bar, 10 μm. d, e Graphs show the number of lysosomes/cell and the percentage of cells with enlarged lysosomes (> 2 μm²) respectively, displayed as mean ± SEM (n = 6). For all graphs *P < 0.05, **P < 0.01, ns = not significant