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. 2018 Sep 18;9:379. doi: 10.3389/fgene.2018.00379

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Copyright © 2018 Ohkuni, Pasupala, Peek, Holloway, Sclar, Levy-Myers, Baker, Basrai and Kerscher.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Slx5 reduces the level of chromatin associated Htt103Q aggregates. (A) Htt103Q, but not Htt25Q, associates with chromatin. Whole cell extracts (WCEs) prepared from equal numbers of cells expressing GFP-tagged Htt25Q-NLS or Htt103Q-NLS from a GAL promoter were fractionated into soluble and chromatin fractions. Htt25Q-NLS-GFP or Htt103Q-NLS-GFP levels in each fraction were monitored by western blot analysis with anti-GFP antibody. Tub2 and histone H3 were used as markers for soluble and chromatin fractions, respectively. (B) WCEs prepared from equal numbers of cells expressing Htt103Q-NLS-GFP with (SLX5-CEN) or without (Vector) were fractionated into soluble and chromatin fractions as described in A. Htt103Q-NLS-GFP levels were monitored by western blot analysis with anti-GFP antibody. Three independent transformants were assayed and shown are the results from two of these. (C) Quantification of the 53 kD GFP signals in soluble fraction from 4B. The 53 kD GFP was normalized using Tub2 levels in soluble fraction. The graph represents the mean of three independent clones with SEM. P-value is 0.0317. (D) Quantification of the aggregate GFP signals in chromatin fraction from 4B. The aggregate GFP signal was normalized using H3 levels in chromatin fraction. The graph represents the mean of three independent transformants with SEM. P-value is 0.0262. (E) Quantification of the 53 kD GFP signals in chromatin fraction from 4B. The 53 kD GFP (shorter exposure) was normalized using H3 levels in chromatin fraction. The graph represents the mean of three independent transformants with SEM. P-value is 0.3523.