Fig. 4

Phosphorylated aSyn increases in concert with double strand breaks and correlate with each other in the nuclear compartment. a Example confocal images (63 × objective) from control (Con) and cases of dementia with Lewy bodies (DLB) showing alpha-synuclein phosphorylated at serine 129 (pS129), double strand break (γH2AX) immuno-fluorescence in neurons (NeuN) and non-neuronal cells.. Intra-nuclear yH2AX and ps129 signal were confirmed via b) Z-stack reconstructed orthogonal views of digitally zoomed neuronal nuclei images captures from DLB cases. Quantification of c) nuclear pS129 (pS129 aSyn^Nuc) and d) γH2AX fluorescence from Con (n = 13 cases) and DLB (n = 12 cases) are presented. ei-ii Frequency distribution of ps129 (per nuclei) are shown as well as fi-ii) correlative levels of ps129 aSyn^Nuc with levels of γH2AX, per case, with spearman's correlation (r) reported. Measures are reported for neuronal (NeuN +) and non-neuronal (NeuN-) populations. Data are expressed in scatterplots with mean ± SEM (in c, d) and as percentage frequency of immunofluorescence intensity per bins of 10 units width (in e). Statistical outcome of Mann–Whitney non-parametric tests and Spearman’s (r) correlation shown in appropriate plots, * = p < 0.05, ** = p < 0.01 and ** = p < 0.001. Scale = 20 µm