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Fig. 3. Treatment with autophagy pathway modulators altered the number of EGFP-fused particles present in cells expressing human ataxin-3 with 84 glutamines. (A) Representative scatterplots reveal a scarcity of EGFP-fused particles in untransfected control cells and altered numbers of particles following treatment with autophagy modulators (3MA and calpeptin). (B) Quantification of the number of DAPI+ nuclei revealed that a similar number of nuclei were present across vehicle-treated and 3MA-treated samples. (C) Comparison of cells treated with vehicle or 3MA (5 mM) revealed a statistically significant increase in detergent-insoluble particles relative to the number of nuclei following 3MA treatment. (D) Similar numbers of nuclei were quantified in samples treated with a vehicle control or calpeptin. (E) Calpeptin treatment produced reductions in detergent-insoluble particles, as detected by FloIT. Data are mean±s.e.m. n=2-6 experimental replicates. *P<0.05, **P<0.01 (one-way ANOVA with Tukey's post-hoc test or Student's t-test, paired, two-tailed). AU, arbitrary units; RFU, relative fluorescence units.