Additional file 5 of Microglial inclusions and neurofilament light chain release follow neuronal α-synuclein lesions in long-term brain slice cultures

Additional file 5: Supplementary Fig. 5. Treatment with highly concentrated αS pff is neurotoxic to Thy1-h[A53T]αS tg HSCs. (A) Immunostaining of pS129-positive aggregates in 50 μm-horizontal sections from Thy1-h[A53T]αS tg HSCs treated with 350 μM αS pff at 1, 2 and 5 weeks post-seeding. Scale bar = 500 μm. (B) Measurement of culture thickness (in μm at the time of fixation) in untreated, 350 μM αS pff-treated and 35 μM αS pff-treated tg HSCs. Mean ± SEM; n(untr., 1 week) = 6, n(untr., 3 weeks) = 18, n(untr., 5 weeks) = 19, n(35 μM pff, 1 week) = 15, n(35 μM pff, 2 weeks) = 10, n(35 μM pff, 3 weeks) = 31, n(35 μM pff, 4 weeks) = 9, n(35 μM pff, 5 weeks) = 18, n(350 μM pff, 1–4 weeks) = 6 each, n(350 μM pff, 5 weeks) = 35 HSCs per group. Note that culture thickness was routinely measured and results come from many different experiments, which accounts for the differences among the n/group. (C) Violin graphs displaying thickness of tg HSCs across different treatments at 5 weeks post-treatment. n(tg, untr.) = 19, n(tg, tg brain hom.) = 24, n(tg, 35 μM pff) = 18, n(tg, 350 μM pff) = 35. Kruskal-Wallis test (H = 45.82; p < 0.0001): **p < 0.0001 Dunn’s multiple comparisons against untreated HSCs. (D) Violin graphs displaying thickness of wt HSCs across different treatments at 5 weeks post-treatment. n(wt, untr.) = 21, n(wt, tg brain hom.) = 27, n(wt, 35 μM pff) = 3, n(wt, 350 μM pff) = 18. Kruskal-Wallis test (H = 11.64; p = 0.0087): **p < 0.01 Dunn’s multiple comparisons against untreated HSCs.