Fig. 2.

Response of control and PD midbrain cells following TLR2 activation. Midbrain cells were treated with 1 μg/ml Pam3CSK4 for 2 and 7 days as indicated. a Cells were stained for P62 (green) and MAP2 (red). Confocal images were taken at 60 × magnification with 8 images captured per condition. Scale bar, 10 μm. b Graph shows fold change in P62 intensity/cell normalised to the control, displayed as mean ± SEM  (n = 6). c Cells were stained for P62 (green) and S100β (red). Confocal images were taken at 60 × magnification with 8 images captured per condition.Scale bar, 10 μm. d Graph shows fold change in P62 intensity/astrocyte normalised to the control, displayed as mean ± SEM (n = 6). e Cells were stained with Lysotracker (red) for 1 h at 37 °C prior to fixation and co-stained for S100β (green). Confocal images taken at 60 × magnification with 8 images captured per condition. Scale bar, 10 μm. f, g Graphs show the average number of lysosomes/astrocyte and the average percentage of astrocytes with enlarged lysosomes (> 2 μm²) ± SEM (n = 6). h Cells were stained for P62 (green) and TH (red). Confocal images were taken at 60 × magnification with 8 images captured per condition. Scale bar, 10 μm. i Graph shows fold change in P62 intensity/TH neuron normalised to the control, displayed mean ± SEM (n = 6). j Cells were stained for α-syn (green) and TH (red). Confocal images were taken at 40 × magnification with 6 images captured per condition. Scale bar, 20 μm. k Graph shows fold change in α-syn integrated density/TH neuron normalised to the control, displayed as mean ± SEM (n = 3). l, m Graphs show flow cytometry analysis of TLR2 in the different cell populations of PD midbrain cells, and the fold change in TLR2 following 24 h Pam3CSK4 treatment, respectfully, displayed as mean ± SEM (n = 3). For all graphs *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001, ns = not significant