FIGURE 7.
Analysis of Yju2 and Prp2 in the spliceosome precipitated with streptavidin Sepharose. (A) Spliceosomes formed with nonbiotinylated (lane 1) or biotinylated pre-mRNA were precipitated with streptavidin Sepharose using extracts prepared from wild-type (lane 2), Ntc90-depleted (lane 3) or NTC30- and NTC20-deleted cells (lane 4), and probed with antibodies against Yju2, Snu114 and Smd1. (B) Spliceosomes formed in the presence of recombinant V5-tagged prp2S387L were precipitated with anti-Ntc20 or anti-V5 antibody in mock-depleted (lanes 1–4), NTC-depleted (lanes 5–8) extracts, or extracts prepared from in vivo Ntc90-depleted cells (lanes 9–12).