Fig. 5. Analysis of RNA splicing in mog-6 mutants. (A) RT-PCR. Genomic DNA (D) or polyA-enriched RNA (A⁺) from wild-type worms were used as positive controls for PCR with sets of oligonucleotides specific for either fem-3 (lanes 1 to 6), fbf-2 (lanes 7 to 12), nos-3 (lanes 13 to 18) or ceh-13 (lanes 19 to 24). Total RNA from either mog-6 or wild-type adults (wt) was used for PCR without (–) or with (+) RT. PCR on genomic DNA was used as a reference for the size of unspliced RNAs (lanes 1, 7, 13 and 19). (B) Northern analysis. Total RNA, 12.5 µg, derived from adult mog-6 or wild-type worms were run on a denaturing agarose gel and probed for mog-6, fbf, nos-3, ges-1 and actin. (C) gld-3 transcripts were analyzed on a separate blot loaded with 25 µg and 50 µg of total RNA from wild-type and mog-6 mutants, respectively. The two major gld-3 transcripts are indicated (Small and Large). Blots were also overexposed to ensure that no minor transcripts of abnormal size were present (not shown).