FIGURE 7.

Spliceosome recycling assays. (A) A scheme for spliceosome recycling assays. (B) Splicing reactions were carried out in wild-type (lanes 1–7) or Δ25 extracts supplemented with affinity-purified NTC (∼5 nM) (lanes 9–15) for 30 min using 0.4 nM labeled actin pre-mRNA. Presplicing was carried out without (lanes 1,8,9) or with fivefold (2 nM), 10-fold (4 nM), or 15-fold (6 nM) excessive amounts of unlabeled wild-type (lanes 2–4,10–12) or ACAC (lanes 5–7,13–15) over labeled pre-mRNA for 30 min prior to the addition of labeled pre-mRNA. (WT) Wild type, (N) no presplicing.