sacy-1(G533R) increases proximal splice site usage of developmentally regulated alt. 3′ splice sites. (A) BLASTp alignment of C. elegans SACY-1 versus human DDX41 at the region around SACY-1(G533). (B) Flowchart of the experimental procedure. The hatched portion is the sequence for which inclusion requires proximal splice site choice. (C) Splicing assay of RNA from synchronized worms. RT-PCR products were run on 6% polyacrylamide denaturing gels. The strain and stage of RNA extraction are shown above each lane. The sequence of the alt. spliced 3′ss is shown to the right of each gel. Quantification and standard deviation are shown below (see Materials and Methods). Each sample shown is representative of three biological replicates; replicates are RNA extracted from independent worm samples grown under identical conditions.
