Fig. 3: SMCHD1 binding to D4Z4 in somatic cells is independent of LRIF1.
a SMCHD1 and LRIF1 ChIP-qPCR in different control32U knock-out conditions. Schematic of one D4Z4 unit and the position of three regions within D4Z4 examined by ChIP-qPCR is indicated. Bars and whiskers represent mean ± SEM of three independent clones. Isotype specific IgG was used for background control. Statistical significance between WT and KO groups was calculated by one-way ANOVA with Dunnett’s post hoc test (**p < 0.01, *p < 0.05). Only significant p-values are shown. b Schematic representation of splicing of the mutant SMCHD1 allele carrying the intronic SNP variant indicated in the box. c SMCHD1 ChIP-qPCR of two D4Z4 regions (DR1 and Q) from four SMCHD1 intron unedited and four SMCHD1 intron edited clones that restores WT SMCHD1 splicing. Bars and whiskers represent mean ± SEM. d LRIF1 ChIP-qPCR of two D4Z4 regions (DR1 and Q) from four SMCHD1 intron unedited and four SMCHD1 intron edited clones. Bars and whiskers represent mean ± SEM. Statistical significance was calculated with unpaired t-test (**p < 0.01, *p < 0.05, ns not significant). e H3 ChIP-qPCR of the D4Z4 DR1 and Q region from four SMCHD1 unedited and four SMCHD1 intron edited clones. Bars and whiskers represent mean ± SEM. Isotype specific IgG was used for background control. f H3K9me3 ChIP-qPCR of the D4Z4 DR1 and Q region from four SMCHD1 unedited and four SMCHD1 intron edited clones. Bars and whiskers represent mean ± SEM. Isotype specific IgG was used for background control. g H3K27me3 ChIP-qPCR of the D4Z4 DR1 region from four SMCHD1 unedited and four SMCHD1 intron edited clones. Bars and whiskers represent mean ± SEM. Isotype specific IgG was used for background control. Statistical significance was calculated with an unpaired t-test (ns not significant).
