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. 2010 Sep 9;6(9):e1001092. doi: 10.1371/journal.ppat.1001092

Figure 6. Assorted CHPKs prevent nuclear and/or cytoplasmic aggresome formation.

Figure 6

(A) U-2 OS cells grown on coverslips were transfected with expression plasmids for Flag epitope tagged Ataxin-Q82 (a spontaneously aggregating protein) as well as the indicated CHPKs (1–8) or an empty vector (EV). Coverslips were harvested 24 hpt and stained for indirect immunofluorescence with antibodies against the Flag epitope, the HA epitope, and Hoechst. Representative images showing Ataxin-Q82 (green), the CHPK (red) and counterstained nuclei (blue) are displayed. (B) Using the same coverslips as in A, the number of cells that contained nuclear aggresomes was determined by counting between 200 and 300 Ataxin-Q82 positive cells in three independent experiments. The percentage of kinase and Ataxin-Q82 positive cells that formed nuclear aggresomes is shown with standard deviation. Asterisks denote statistically significant differences from the empty vector control (* indicates P≤0.05, ** indicates P≤0.01). (C) Cytoplasmic aggregates formed by the Ataxin-Q82-K772T were imaged as in (A) above. (D) Cytoplasmic aggresomes were quantitated as in B above except that four independent experiments were analyzed.