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. 2006 May;12(5):862–871. doi: 10.1261/rna.2319806

FIGURE 1.

FIGURE 1.

Spliceosome assembly and activation in snu114–60 and snu114–12 extracts. (A) SNU114 and snu114–60 extracts were incubated at 23°C and 30°C with biotinylated pre-mRNA and spliceosomes were affinity-purified. Levels of snRNAs associated with transcript after 20 min of incubation under splicing conditions are shown, as quantified by real-time PCR. Each coprecipitating snRNA was normalized to 1.0 for SNU114 at 23°C. As a control, SNU114 extract was incubated at 23°C with pre-mRNA in the absence of ATP. (B) Ratio of transcript-bound U4 snRNA vs. U6 snRNA over time for SNU114 and snu114–60 extracts. (C) snRNAs bound to biotinylated transcript after 20 min of incubation under splicing conditions in SNU114 and snu114–12 extract at 30°C. Each snRNA was normalized to 1.0 for SNU114. (D) Ratio of transcript-bound U4 snRNA vs. U6 snRNA over time for SNU114 and snu114–12 extracts. Data represent the average of at least five experiments; in A and B, n = 7 for the 30°C data and n = 5 for the 25°C data, and in C and D, n = 5. Error bars indicate standard error.