Fig. 5: Loss of imprinting at Slc38a4 upon maternal transmission of the MT2A KO allele.
From: Evolution of imprinting via lineage-specific insertion of retroviral promoters
a Genome-browser screenshot of the mouse Slc38a4 promoter and upstream region, including the MT2A LTR (red), annotated Slc38a4 exon 1, CGI (green), and igDMR (blue). GVO RNA-seq as well as RNA pol II, H3K4me3 and H3K36me3 ChIP-seq tracks are shown, along with DNAme data for GVO, sperm and adult liver. The region within the igDMR analyzed by sodium bisulfite sequencing (SBS), which includes 11 CpG sites, is shown at the bottom. Δ: extent of the MT2AKO deletion allele. b DNAme of the Slc38a4 igDMR in GVO from wild-type and Slc38a4MT2AKO/MT2AKO females determined by SBS. c DNAme of the Slc38a4 igDMR in E13.5 (Slc38a4+/MT2AKO × CAST)F1 embryos determined by SBS. Data for control (+/+C) and heterozygous (KO/+C) littermates with a maternally inherited MT2AKO are shown. +C: wild-type CAST allele; KO: Slc38a4MT2AKO. A polymorphic insertion in the amplified region allows for discrimination of maternal (Mat) and paternal (Pat) strands. Allele-specific expression analyses of F1 E13.5 placental RNA by d RT-PCR followed by PvuII RFLP analysis (RT reverse transcriptase), and e, f Sanger sequencing of a T ⟷ C transition in the 3′UTR of the Slc38a4 cDNA (maternal B6: T allele; and paternal CAST: C allele). Each bar in f shows the mean of individual samples, and error bars show S.D. of two SNPs analyzed. Source data are provided as a Source Data file. g Total Slc38a4 mRNA levels in E8.5, E13.5, E16.5, and E18.5 placentae, as determined by RT-qPCR (n = 6 biologically independent samples for each datapoint). Expression levels are relative to the housekeeping gene Ppia. Graph shows mean ± S.D. Source data are provided as a Source Data file.
