Fig. 2: EHMT1 suppresses expression of miRNA.

A ChIP-qRT-PCR analysis of H3K9me2 modification within 3 regions of NRSF/REST promoter (P1, P2, P3) and surrounding (P1, P2) and distal to (P3) the Translational Start Sites (TSSs) of miR-142, miR-153 and miR-26a promoters of pluripotent hiPSC (see Fig. S3). ChIP was performed with an anti-H3K9me2 antibody, and H3K9me2 enrichments were analysed by qRT-PCR. Enrichment is plotted as increase relative to the input DNA in specific genomic regions in the absence of ChIP antibody primer sets are listed in Table S4. B–D Validation of miRNA-seq data by qRT-PCR to confirm miRNA-seq results: (B) UNC0638-treated hiPSC; (C) KS-patient iPSC; (D) UNC0638-treated mECS. The qRT-PCR data were shown as Mean±SEM and analysed by One-way ANOVA with post hoc comparisons using Dunnett’s multiple comparisons test comparing to control samples. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n ≥ 3. E Relative mRNA abundance measured by qRT-PCR of the REST-target genes ACTA1, NRXN3, and Calbindin1 (CALB1) in pluripotent hiPSCs treated with UNC0638. Induction of RESTΔUTR with doxycycline (DOX) suppresses the UNC0638-induced gene expression. Results are plotted as % of mRNA abundance in the absence of DOX, shown as dotted line. F, i Venn diagrams showing association between miRNAs following UNC0638 treatment and those associated with Intellectual Disability (ID) and schizophrenia (SCZ). Crossover probability was assessed for each disorder by calculating hypergeometric probability, with P < 0.05 considered statistically significant; (ii) Venn diagram to show overlap of REST targeting miRNAs between ID, SCZ and ASD.