Development of Small Molecular Proteasome Inhibitors Using a Caenorhabditis elegans Screen

Figure 3

Screen using rrf-1(pk1417)(I);ozIs2(II). (a) Paired DAPI (nuclei, left) and GLD-1::GFP (right) images from animals treated with carrier control (A) or compound 73T (B, C, D). Representative images of animals scored positive for GLD-1 misexpression are shown in panels B–D and quantified on the graph (bottom). The primary phenotypes associated with GLD-1::GFP were a transient increase in GLD-1::GFP (B), extension of into oocytes (C), and abnormal nuclear morphology coupled with ectopic GLD-1::GFP expression (D). For all images the “*” = distal tip of the germ line (mitosis), dashed line is at the loop region (pachytene-diplotene/diakinesis boundary), curved line ending in an arrow follows the germ line, closed arrow heads are examples of ectopic GLD-1::GFP expression, and “” indicates abnormal nuclei. All GFP exposure times were identical for control and test conditions and all images are from 6 hours of treatment. (b) Compounds 73T, 53P, 33K, 5B, clasto-lactacystin β-lactone, and epoxomicin were tested for their ability to cause the expression of GLD-1::GFP. Gonad arms with changes in GLD-1 expression were graphed as a faction of total arms scored (%) for each concentration. To illustrate the dispersion of the data, the bars indicated ± SD (standard deviation) of samples for each point.

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