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. 2020 Feb 3;11(2):84. doi: 10.1038/s41419-020-2285-7

Fig. 6. αSyn fragments that induce intracellular aggregation also induce toxicity.

Fig. 6

a Experimental design. Cells were differentiated for 4 days before start of treatment. Treatments with recombinant FL-αSyn and fragments were carried out for 48 h. Toxicity was assessed at three readout times. b–d Toxicity measured by LDH release in cells treated with recombinant FL-αSyn and fragments at DIV6 (b), DIV9 (c) and DIV12 (d). At DIV12, recombinant αSyn fragments 61–140 and 1–95 induced toxicity levels of approximately 30% compared to a cell lysis positive control. Data are presented as mean + SEM from 4 biological repeats with at least 4 technical replicates each. ns: not significant, ***p < 0.001; one-way ANOVA with Tukey’s post hoc test. e, f Percentage of condensed nuclei in cells treated with recombinant FL-αSyn and fragments at DIV12. Cells were fixed and nuclei were stained with DAPI. Recombinant αSyn fragments 61–140 and 1–95 induced higher toxicity levels in accordance with LDH data (d). Data are presented as mean + SEM from 3 biological replicates with at least 3 technical repeats each and 3 pictures per technical repeat. ***p < 0.001; one-way ANOVA with Tukey’s post hoc test. f Representative pictures of DAPI stained nuclei from cells treated with recombinant FL-αSyn and fragments at DIV12. Red arrowheads indicate condensed nuclei. Scale bar: 30 µm.