FIG. 2.

Ssl38 is required for efficient splicing. (A) Ssl38 coimmunoprecipitates with Prp1. Soluble protein extracts were subjected to immunoprecipitation with anti-GFP or anti-myc antibodies. Inputs (lanes I), immunoprecipitated (lanes IP) fractions, and proteins washed away (lanes FT) were analyzed by Western blotting with anti-myc or anti-GFP antibodies. (B) Unspliced transcripts accumulate in ssl38-ts. The scheme of the RT-PCR assay used to assess splicing efficiency is shown. Total RNA extracted from indicated cells was reverse transcribed in the presence (+RT) or absence (−RT) of reverse transcriptase. Complementary DNAs (cDNAs) were PCR amplified and in-gel quantified. Unspliced, upper band; spliced, lower band. (C) The steady-state level of act1 transcripts or 25S RNA is not overtly affected by ssl38-ts. Same procedure as in panel B, except that cDNAs were quantified by real-time PCR. The indicated values correspond to averages and mean deviations (m.d.) calculated from three (25S RNA) or two (act1) independent samples.