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. 2016 Aug 11;6:31393. doi: 10.1038/srep31393

Figure 8. Effect of mutations in the distal 3′ SL of U6atac snRNA on U12-dependent splicing in vivo.

Figure 8

(a) Sequence of the WT U6atac distal 3′ SL and mutations made in the SL are illustrated. These mutations were made in the U6atac snRNA carrying the GG14/15CC first site mutation. (b) Splicing phenotypes of the P120 WT and the P120 CC5/6GG + UC84/85AG mutant coexpressed with the indicated suppressor snRNA constructs. CHO cells were transiently transfected with the indicated constructs and total RNA was extracted. The splicing pattern of the U12-dependent P120 intron was analyzed by RT-PCR using primers designed to bind flanking exons. Lane M: 100 bp ladder, lane EV: Empty vector; RT(-), without reverse transcriptase. The positions of bands corresponding to unspliced RNA, RNA spliced at the normal U12-dependent splice sites (U12 spliced) and RNA spliced at the cryptic U2-dependent splice sites (U2 cryptic) are indicated. (c) Quantitative analysis of the U12 unspliced/spliced bands. Numbers (x-axis) correspond to the respective lanes of the gel shown in (b). Error bars represent ± SE of three experiments.