Development of Small Molecular Proteasome Inhibitors Using a Caenorhabditis elegans Screen

Figure 2

The rrf-1(pk1417)(I);ozIs2(II) (GLD-1::GFP) strain can be used to score GLD-1 levels in real-time. (a) DIC (differential interference contrast), GFP, and merged images of a live animal expressing GLD-1::GFP. The distal tip of the germ line is indicated by the “*” and the dashed line is at the loop region (pachytene-diplotene/diakinesis boundary). (b) The expression levels of GLD-1::GFP are similar to endogenous GLD-1. Western blotting was performed on lysates of 25 and 50 worms of N2 ((A), wild-type), rrf-1(pk1417)(I);ozIs2(II) (B), and gld-1(q485)(I);ozIs2(II) (C). GLD-1 and GLD-1::GFP are indicated by the arrows. N2 has a single band at approximately 60 kDa (A). As expected, a strain carrying the ozIs2 transgene in the gld-1 null background only has the larger GLD-1::GFP fusion band at approximately 85 kDa (B). The rrf-1(pk1417)(I);ozIs2(II) has both bands from endogenous GLD-1 and the GLD-1::GFP fusion (C).

(a)

(b)