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. 2012 Dec 7;41(2):1294–1306. doi: 10.1093/nar/gks1188

Figure 5.

Figure 5.

UAP56 associates with the pre-mRNA independent of Aly or THO. (A) Ftz pre-mRNA was incubated in HeLa nuclear extract under splicing condition for 30, 60, 90 or 120 min, followed by mRNA IPs using indicated antibodies; 25% of the input was loaded. Quantification of three independent experiments is shown below. In the left graph, bars indicate the relative pre-mRNA IP efficiency calculated by dividing the IP efficiency of pre-mRNA at each time point by that of spliced mRNA at 120 min. In the right graph, bars indicate the ratio of IP efficiency for the pre-mRNA to that for the spliced mRNA. Error bars represent standard deviations (n = 3). (B–D) Ftz pre-mRNA was incubated under splicing condition in mock (B), ΔAly (C) or ΔTHO (D) nuclear extract, respectively, for 30, 90 or 120 min followed by IPs using indicated antibodies; 25% of the input was loaded. (E) Bars indicate relative pre-mRNA IP efficiencies of U2AF35 at each time point that are calculated by dividing pre-mRNA IP efficiencies in ΔAly and ΔTHO extracts by those in Mock extract. Error bars represent standard deviations (n = 3). (F–H) Similar to (E), except that pre-mRNA IP efficiencies of the UAP56 antibody (F), spliced mRNA IP efficiencies of the U2AF35 antibody (G) and spliced mRNA IP efficiencies of the UAP56 antibody are indicated.