Figure 3.

Trans-splicing requirement for an ESE and SF2/ASF. (A) Control AdML PAR pre-mRNA cis-splicing was carried out in nuclear extract (NE) or in S100 extract complemented with SF2/ASF. Lane 1, size markers. (B) Trans-splicing reactions were performed in nuclear extract or in S100 extract with or without SF2/ASF. ³²P-labeled AdML ΔAG was incubated for 1 hr, and then excess ³H-labeled M2-SF2W, or M2-Δ, or water was added for the second hour. Trans-spliced mRNA (TS, lanes 4 and 6) is marked by a black arrow, and the expected position of the M2-Δ mRNA is marked with a dashed open arrow. (C) A reciprocal labeling experiment is shown. The top part of the autoradiogram is a longer exposure.