Fig. 4: Gtl2 polyA knock-in results in dysregulation of placental genes in the Dlk1-Dio3 imprinted domain at E12.5.

a RNA-seq demonstrates mRNA expression in Gtl2 polyA knock-in and WT placentas overall chr12. Dlk1-Dio3 imprinted domain is annotated in gray shade. b Compare gene expression of Dlk1, Rtl1 and Dio3 in the Dlk1-Dio3 domain among Gtl2 polyA knock-in and WT placentas using RNA-seq data. c qRT-PCR analysis confirm the expression of Dlk1, Rtl1 and Dio3 to 18 s in Gtl2 polyA knock-in placentas. d RNA-seq demonstrates lncRNA expression in Gtl2 polyA knock-in and WT placentas overall chr12. Dlk1-Dio3 domain is annotated in gray shade. e Compared gene expression of Gtl2, Rian, and Mirg in the Dlk1-Dio3 domain among Gtl2 polyA knock-in and WT placentas using RNA-seq data. f qRT-PCR analysis confirm the expression of Gtl2, Rian, and Mirg to 18 s in Gtl2 polyA knock-in placentas. g Small RNA-seq demonstrates miRNA expression in Gtl2 polyA knock-in and WT placentas overall chr12. Dlk1-Dio3 domain is annotated in gray shade. h Compare expression of miRNAs in Dlk1-Dio3 domain detected by small RNA-seq in four genotype placentas. Seven miRNAs shown in light blue shade are located in Gtl2 locus, five miRNAs shown in light pink shade are located in Rtl1 antisense locus, three miRNAs shown in light green shade are located in Rian locus, and twenty-six miRNAs showed in light yellow shade are located in Mirg locus. The miRNAs shown in purple have been detected by qRT-PCR. i qRT-PCR analysis confirm the expression of miR-337, miR-127, miR-370, miR-381 and miR-541, which we select from each locus in Dlk1-Dio3 domain whose expression abolish in MKI and HOMO placentas but are normal in PKI placentas. In (c, f, i), biological duplication: WT n = 6, PKI n = 6, MKI n = 6, HOMO n = 4. The mean expression ± SD of each genotype is plotted. Student t-test is used to analyze the p values. *p < 0.05, **p < 0.01. In (b, c, e, f, h, i), gene expression levels in WT are arbitrarily set to 1.