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. 2006 Sep;12(9):1738–1746. doi: 10.1261/rna.120606

FIGURE 4.

FIGURE 4.

Spliceosomal A complexes purified from U4/U6-depleted nuclear extract are functional. (A) RNA composition of spliceosomal complexes purified from U4/U6-depleted nuclear extract. RNA was analyzed as in Figure 2A and visualized by silver staining (upper panel) or by autoradiography (lower panel). (B) Splicing activity of A complexes purified from U4/U6-depleted extract. Purified spliceosomal A complexes (lanes 1,2) or an equimolar mixture of the purified A complex and naked, untagged MINX pre-mRNA (lanes 3,4) were incubated in the presence of U2-depleted extract for 3 h at 0°C (lane 1) or at 30°C (lanes 2,3), or for 3 h at 30°C in U2-depleted extract complemented with 12S U2 snRNPs (lane 4). RNA was analyzed as in Figure 2B.