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. 2010 Nov 1;24(21):2389–2394. doi: 10.1101/gad.1974810

Figure 3.

Figure 3.

Urp is required for the second step of U2-type splicing. (A) Conventional splicing assay. Urp or, as a control, U2AF35 was analyzed for its ability to complement splicing of the Ad ML pre-mRNA substrate in the ΔUrpNE. Splicing of Ad ML in NE (md) is shown as a control. (B) Splicing complex assembly. Urp was analyzed for its ability to support assembly of the prespliceosome (complex A) and mature spliceosome (complexes B and C) on the Ad ML pre-RNA substrate in the ΔUrpNE. (C) Bimolecular ligation assay. Uniformly labeled 5′ RNA substrate was preincubated in the ΔUrpNE for 0 or 1 h. After 1 h of incubation, an excess of unlabeled 3′ RNA substrate was added in the presence or absence of Urp, followed by further incubation for 1 h. Samples were analyzed by denaturing PAGE.