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Analysis of snRNAs in purified spliceosomes. Spliceosomes were purified using AdML-M3, AdML, Ftz-M3, or Ftz pre-mRNAs as substrates (AdML or Ftz pre-mRNAs lacking hairpins were used for negative controls). Total RNAs extracted from the nuclear extract (NE), AdML, AdML-M3, Ftz, or Ftz-M3 elutions were 32P-end-labeled (+) or not labeled (−) and fractionated on an 8% polyacrylamide gel. Note that the snRNAs are 3′-end-labeled differentially with RNA ligase (15).
