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. Author manuscript; available in PMC: 2009 Apr 14.
Published in final edited form as: Nat Struct Mol Biol. 2008 Sep 14;15(10):1040–1048. doi: 10.1038/nsmb.1485

Figure 3.

Figure 3

SRp38 promotes formation of spliceosomal complex A. (a) Spliceosome-assembly assays were carried out in S100 complemented with the indicated components and the β-SRp38 pre-mRNA. Splicing complexes were resolved on a 1.5% low-melting agarose gel. (b) SRp38 activates AdML-SRp38 in in vitro splicing assays. Splicing was performed in S100 supplemented with 50 ng of GST-tagged SRp38 with (lane 1) or without (lane 3) NF40–60. Products of splicing were analyzed by denaturing PAGE and autoradiography. (c) Spliceosome assembly was performed the same as in a, except with the AdML-SRp38 pre-mRNA. NE, nuclear extract. (d) Spliceosome-assembly assays were carried out as in c, except with the additions indicated at the top. Anti-U1, anti-U2, anti-U5 or anti-U6 snRNA oligonucleotides (5 µM) were added to reaction mixtures. Endogenous ATP was depleted by preincubating S100 at 30 °C for 40 min.