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

Figure 4.

Figure 4.

Aly and THO are required for UAP56–CBC interaction and stable association of TREX components with the spliced mRNA. (A) IPs were performed from RNased Mock, ΔUAP56, ΔAly and ΔTHO extracts as well as ΔAly extract supplemented with purified His-Aly using indicated antibodies. Western analyses were performed with the CBP80 antibody. (B) Schematic of the interaction between CBC and TREX. Aly and THO interact with CBP80 and mediate the interaction between TREX and CBC. (C) AdML pre-mRNA was incubated in Mock or ΔAly nuclear extract under splicing condition followed by mRNA IPs using indicated antibodies. The control antibody was an antibody against fSAP130; 25% of the input was loaded. Quantifications of three independent experiments are shown in the right panel. The bars indicate the ratio of IP efficiency in ΔAly extract relative to the corresponding IP efficiency in Mock extract. Error bars represent standard deviations (n = 3). (D) Same as (C), except that ΔTHO nuclear extract was used instead of ΔAly nuclear extract. (E) ΔAly nuclear extracts supplemented with purified recombinant His-Aly or His-AlyΔC proteins were used for mRNA IPs with indicated antibodies.