FIGURE 8.

No evidence for intronic polyadenylation in intron 6 of IL7R. 3′ RACE was used to investigate intronic polyadenylated isoforms of IL7R in total RNA from Jurkat cells that naturally express IL7R. (A) Schematic of the IL7R pre-mRNA. RT reactions were performed with an Oligo(dT)-Anchor primer either in the presence or absence of RT enzyme. The first round of PCR was performed with a forward primer in exon 3 and a reverse primer that anneals to the anchor sequence incorporated by the Oligo(dT)-Anchor primer during the RT step. This was followed by nested PCR reactions using the primers indicated. (B, top) A diagram of the expected PCR products with primer pairs indicated in lower panels. (B, bottom) Nested PCR products using the indicated primer pairs were resolved on 5% native polyacrylamide gels and exposed to PhosphorImager screens for visualization.