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. 2011 Mar 1;25(5):440–444. doi: 10.1101/gad.2009411

Figure 2.

Figure 2.

E7 results in formation of a labile spliceosome and blocks the A complex assembly. (A) CMV-Ftz was incubated in coupled transcription/splicing mixtures in the presence of E7 or PlaF for 8 min. CMV-Ftz was omitted in lane 3. Heparin (0.5 μL) was added to the reaction mixtures, and then an aliquot was fractionated on a 1.2% native low-melting agarose gel and detected by PhosphorImager. The spliceosome (ssome) is indicated. (B) Same as A, except using CMV-Ftz and CMV-Δss DNA templates. The spliceosome and nonspecific Δss complexes are indicated. (C) Same as A, except 0.5 μL and 1.5 μL of heparin was added. The white box to the right of lane 4 indicates the heparin-sensitive E7 spliceosome. (D) T7-AdML was incubated under standard splicing conditions in E7 or no drug for the indicated times, heparin was added, and spliceosomal complexes were separated on a 1.5% native low-melting agarose gel.