Fig. 6: Allelic expression analysis of the tammar MESTIT1 and CEP41B.

A Isoforms of MESTIT1 and primers for allelic expression analysis. A SNP site was found in the common region of the two isoforms (arrowhead). In the shorter isoform, specific primers could not be designed because the SNP site was close to the poly-A tail. However, primers used for the SNP search could detect the longer isoform. B Allelic expression of the longer isoform of MESTIT1 in the tammar placentas. The imprinting status of the tammar MESTIT1 was determined by direct sequencing of PCR products that contained a SNP site. In contrast to the genomic DNA data in which double peaks at the SNP site have almost the same signal strength, the cDNA data clearly showed that the lncRNA is predominantly expressed from either one of the two alleles. C Exon structures of the two CEP41 isoforms. Orange and aqua coloured boxes represent the CEP41B-specific and CEP41A-specific exons, respectively. Blue arrows represent primers used for detecting the genomic SNP site. D Primer design for the allelic expression analysis of the CEP41B transcript. Allelic expression analysis of CEP41B was performed using CEP41B specific forward primer and a reverse primer designed for the 3′ UTR common amongst CEP41 isoforms. Arrowheads represent SNP sites. E Allelic expression of the tammar CEP41B by direct sequencing with PCR amplification. In animal #1, skewed maternal expression was observed in the placenta tissues. However, in animal #2, skewed paternal expression was observed in the TOM tissue. Due to low expression levels, it was not possible to determine allelic expression of CEP41B in the BOM of Animal #2.