Figure 5.

mCherry::ubiquitin colocalization to Q82::GFP aggregates. Worms co-expressing an mCherry::ubiquitin with Q82::GFP were exposed to RNAi of ubc-22, ubc-1, or pL4440 control. A typical control aggregate is shown in (A) with a 10.16-micron line (45 pixels) measuring a cross-section of fluorescence intensity. Data for this line profile is shown in (B). (C) Averaged line profile data for mCherry::ubiquitin fluorescence of Q82::GFP aggregates at multiple time points. Higher levels of colocalization of mCherry::ubiqutin occur at an age of 36 hours. Peaks represent fluorescence values in the aggregates while the peripheral points represent intracellular fluorescence. Values are the mean (± SEM) of all values at that distance coordinate for aggregates at the given time point. All values have been normalized with respect to exposure time and background levels of fluorescence. (D) Time course of mCherry::ubiquitin colocalization to Q82::GFP aggregates shows the spike in ubiquitin colocalization at 36 hours, which is diminished by RNAi of ubc-1 or ubc-22. Data shown are the mean (± SEM) ratio of mCherry fluorescence to GFP fluorescence from line profile measurements of composite images for multiple aggregates at each time point.