Extended Data Fig. 10: Increased methylation levels at the Line 1 5’ region and the Gck promoter and impaired glucose tolerance in the offspring from oocyte-specific Tet3-conditional gene knockout dams.
From: Maternal inheritance of glucose intolerance via oocyte TET3 insufficiency
a–b, Illustrative diagram of experimental groups with reduced levels of maternal TET3 expression induced by maternal hyperglycemia (a) and oocyte-specific Tet3 heterozygous and homozygous knockout (b). c, BS analysis of the H19 imprinted control region in the sperm and male pronuclei isolated from aphidicolin-treated zygotes of PN3–4 stage. d, BS analysis of the Line 1 5’ region in the sperm and male pronuclei from aphidicolin-treated PN3–4 zygotes. e, BS analysis of the Gck promoter region in the female pronuclei isolated from aphidicolin-treated zygotes of PN3–4 stage. c–e, Open and filled circles represent unmethylated and methylated CpG sites, respectively. The percentages of methylated CpGs are indicated. f–g, Pyrosequencing analysis of the Gck promoter in pancreatic islets from offspring at 3 weeks of age (male: f, female: g). n of the group is indicated. P values for comparisons of Mat-Het and Mat-WT are presented using *, Mat-KO and Mat-WT using †. h, Gck mRNA expression in female pancreatic islets at 3 weeks of age. Mat-WT, Mat-Het, Mat-KO: n = 8, 10, 11 mice, respectively. i, Western blot analysis of GCK in pancreatic islets of 3-week-old offspring males. Each sample was pooled islets from 3–5 mice. Gel source data are presented in Supplementary Figure 1. j–k, GTT results of female offspring at 16 weeks of age (j) and the corresponding AUCs (k). Mat-WT, Mat-Het, Mat-KO: n = 11, 13 and 12 mice, respectively. P values for comparisons of Mat-Het and Mat-WT are presented using *, Mat-KO and Mat-WT using †. f–k, Data are presented as the mean ± s.e.m.; two-tailed P values were calculated by one-way repeated-measurements ANOVA followed by the post hoc unpaired t-test with Mat-WT (j) and the unpaired Student’s t-test with Mat-WT (f–h, k). *P < 0.05, **P < 0.01 and ***P < 0.001; ††P < 0.01 and †††P < 0.001. Statistical details are in Supplementary Table 5.
