Fig. 5.

Robinson et al., 2021 - Flow cytometry allows rapid detection of protein aggregates in cellular and zebrafish models of spinocerebellar ataxia 3
Other Figures
All Figure Page
Back to All Figure Page
Fig. 5.

Flow cytometric analysis of dissociated cells from whole zebrafish can be used to rapidly quantify insoluble EGFP ataxin-3 particles and screen compounds for the effect on proteinopathy. (A) Representative scatterplots of flow cytometric populations from dissociated 6-day-old zebrafish revealed changes in the number of detected detergent-insoluble particles despite similar numbers of nuclei. (B,C) Zebrafish expressing human mutant ataxin-3 displayed similar numbers of RedDot-stained nuclei but significantly more detergent-insoluble ataxin-3 particles at 2 days of age. (D) Examination of insoluble particle size revealed similar-sized particles across genotypes. (E,F) At 6 dpf, cells from dissociated zebrafish showed similar numbers of nuclei but more EGFP ataxin-3 particles were detected in cells obtained from zebrafish expressing human mutant ataxin-3 compared to non-transgenic siblings and zebrafish larvae expressing human wild-type ataxin-3. (G) Detergent-insoluble particles were a similar size in cells obtained from zebrafish expressing wild-type human ataxin-3 and mutant human ataxin-3. (H) Treatment with the autophagy inhibitor chloroquine (3 mM) increased the number of detergent-insoluble particles detected. (I) Treatment with the autophagy inducer calpeptin decreased the number of detergent-insoluble particles in 2-dpf zebrafish expressing mutant ataxin-3. Data are meanĀ±s.e.m. n=2-9 experimental replicates. *P<0.05; **P<0.01; ***P<0.001; ns, not significant (one-way ANOVA with Tukey's post-hoc test or Student's t-test, paired, two-tailed). AU, arbitrary units; RFU, relative fluorescence units.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Dis. Model. Mech.