Fig 4. Detection of processed V2 transcripts.
A. Diagram of 32P-labeled antisense RNA probe used for RNase protection assays, with sizes (in nt) of each exon and the intronic region downstream from exon 20A shown below.
B. RNase protection assays. Each lane contains the probe shown in Panel A plus 3 μg of each of the following RNAs: Lane 1, probe alone. Lane 2, tRNA control. Lanes 3 and 4, normal theca cells from two individuals. Lanes 5 and 6, Theca cells from two PCOS individuals. Lane 7 total RNA from NCI-H295A cells. Lane M, 32P-labeled 1 kb DNA ladder (Invitrogen). The additional bands seen with NCI-H295A RNA may represent additional alternatively spliced forms. Sequences of primers used for probe preparation are shown in Table 3.