Fig. 2

G2019S LRRK2 hippocampal neurons show mild, reversible elevation in induced α-synuclein pathology 21 days post-transduction. Primary hippocampal neurons from NTG (a) or G2019S (b) pups were transduced with α-synuclein PFFs and allowed to age a further 21 days prior to fixation and staining for pS129 α-synuclein (magenta), MAP2 (gray) and NeuN (blue). The neurons were additionally treated with LRRK2 inhibitors PF-475 and PF-360 2 days prior to transduction and fed with media containing inhibitors each week thereafter. No large differences can be observed in the type or abundance of α-synuclein pathology. c Quantification of α-synuclein pathology reveals a mild elevation in G2019S neurons, which is reversible with 30 or 120 nM PF-360. *P < 0.05 by Dunnett’s multiple comparison test between NTG and G2019S neurons or Sidak’s multiple comparisons test between G2019S neurons treated with LRRK2 inhibitors. d MAP2 area is reduced with 21 days α-synuclein PFF treatment in both NTG and G2019S neurons, and is not significantly affected by LRRK2 inhibitor treatment. *P < 0.05 by 2-way ANOVA with Dunnett’s multiple comparison test for comparison within genotype. e The number of neurons, as quantified by NeuN number, is reduced with 21 days α-synuclein PFF treatment in both NTG and G2019S neurons, although not significantly by 2-way ANOVA followed by Dunnett’s multiple comparison test and is not significantly affected by LRRK2 inhibitor treatment. (N = 12 biological replicates). Means + s.e.m.; all values are normalized to NTG neurons treated with α-synuclein PFFs and DMSO. Scale bars = 50 μm