Fig. 1: Nestin-Cre mediated loss of Ehmt2 in forebrain is sufficient to unsilenced the imprinted Snrpn gene in the maternal chromosome.
From: Mechanism of EHMT2-mediated genomic imprinting associated with Prader-Willi syndrome
a Schematic figure shows a strategy for generating Ehmt2 cKO mice with maternal Snrpn-EGFP (Exon 3 of Snrpn is fused in-frame with EGFP, Ex3-EGFP). b The level of EHMT2 protein was significantly decreased in forebrains of Ehmt2 cKO (Nestin-Cre + ;f/f;p+/mS-E) mice at p10. c H3K9me2 level was decreased in forebrains of Ehmt2 cKO mice at p10. b, c Data are presented as mean ± SD. (Two-tailed Student’s t test; p = 0.0083 and p = 0.0023, respectively). d RT–PCR analysis detected the expression of Snrpn-EGFP (Ex3-EGFP) in the forebrains of Ehmt2 cKO mice carrying maternal Snrpn-EGFP at p10 (RTase: +/−, with or without reverse transcriptase). e Quantitative RT–qPCR analysis of Snrpn and Snrpn-EGFP mRNA levels in the forebrains at p10. Data are presented as mean ± SEM (Two-tailed Student’s t test; ****p  <  0.0001). f ChIP–qPCR quantification of H3K9me2 and H3K9ac in brains of Ehmt2 cKO mice at E18. Enrichment of H3K9me2 was significantly reduced PWS-IC (IC-1 and IC2) of Ehmt2 cKO mice. Data are presented as mean ± SEM (Two-tailed Student’s t test; p value is indicated in graphs). g Comparison of the DNA methylation in PWS-IC between control and Ehmt2 cKO mice at E18 (Two-tailed Student’s t test; ns, not significant). h The genes in human 15q11.2-q13 and its homologous region mouse central chromosome 7. Created in BioRender. Qiao, S. (2025) https://BioRender.com/v2500u0. The paternally expressed genes are highlighted in blue. Heat map of the expression of genes located in the central chromosome 7 between control (Ehmt2f/f;m+/pSnrpn-EGFP) and Ehmt2 cKO (Nestin-Cre+/−;Ehmt2f/f;mSnrpn-EGFP/p+) forebrain at p10 (adj p value < 0.05). i, j ChIP-qPCR quantification of EHMT2 and H3K9me2 on imprinted genes in control (Ehmt2f/f;mSnrpn-EGFP/p+) and Ehmt2 cKO (Nestin-Cre+/−;Ehmt2f/f;mSnrpn-EGFP/p+) forebrain at p10. Data are presented as mean ± SEM (One-tailed Student’s t test; p value is indicated in graphs). Source data are provided as a Source Data file.
