Minor intron retention and reduced spliced mRNAs from MIGs in miso mutant ovaries. (A) SSun of 5′ and 3′ splice sites of minor introns, as determined by RNA-seq of ovary RNAs. Mean ± SD (n = 3). P-value < 0.05 (Student's t-test, unpaired, two-tailed) is indicated by *. (B) RT-qPCR quantification of RNAs with retained minor introns at 5′ and 3′ splice sites, normalized to act5c mRNA. Mean ± SD (n = 3). P-value < 0.05 (Student's t-test, unpaired, two-tailed) is indicated by *. (C) RT-qPCR quantification of spliced mRNAs from MIGs and gapdh2 in ovaries, normalized to act5c mRNA. Mean ± SD (n = 3). P-value < 0.05 (Student's t-test, unpaired, two-tailed) is indicated by *. (D) Agarose gel electrophoresis of RT-PCR products from ovary RNAs, stained with SYBR Safe. Bands representing spliced and unspliced RNAs are indicated. Results for lsm12a, sf3a1, and stas are shown as representative. The results for the other MIGs and gapdh are shown in Supplemental Figures S4, S6, and S7.
