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FRAP analysis of mCherry::ubiquitin and Q82::GFP in polyglutamine aggregates reveals differential mobility of ubiquitin and polyglutamine proteins within aggregates. Worms co-expressing an mCherry::ubiquitin fusion protein with Q82::GFP were subjected to FRAP using a confocal laser scanning microscope system. Fluorescence intensity is indicated by a heat map of either Q82::GFP (A) or mCherry::ubiquitin (B). Red or yellow rectangles indicate regions to where bleaching was directed. White rectangles indicate regions that were used to control for acquisition photobleaching. Measurements of fluorescence recovery were taken every 0.1 s for 3 minutes. Quantitative analysis (C) of fluorescence recovery in bleached regions indicates higher overall mobility of mCherry::ubiquitin fusion protein when compared to Q82::GFP protein. Data plotted are the mean ± SEM.
In the FRAP experiments, mCherry or GFP was bleached within a
region of the Q82::GFP aggregates in adult worms and recovery was
observed (Figure 6A, B).
Q82::GFP showed a slight recovery (Figure 6C) with a mobile
fraction of 23.3 ± 9.2.
Skibinski, Gregory A; Boyd, LynnJournal: BMC Cell Biology
Issue 1DOI: 10.1186/1471-2121-13-10Published: 2012-12-01Institution(s):
University of Alabama in Huntsville
This image is from the article titled "Ubiquitination is involved in secondary growth, not initial formation of polyglutamine protein aggregates in C. elegans
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