Figure 2
SLY controls the expression of genes involved in chromatin regulation during postmeiotic sperm differentiation. (a) Representation of SLY ChIP-Seq, its input, H3K4me3 ChIP-Seq, Kcr ChIP-Seq, H3K27me3 ChIP-Seq and H3K9me3 ChIP-seq at the TSS of Dot1l, one of the genes found to have the highest enrichment of SLY by ChIP-Seq. (b) Transcript level of autosomal genes found to be highly enriched in SLY protein at their TSS (10% of genes with highest SLY peak, Supplementary Figure 3) and of XY-encoded genes, measured by RT-qPCR in Sly-KD and WT round spermatids. The graph represents the geometric mean ±S.E.M (after normalization with β-actin, n=3–6 samples per genotype). For all except two genes (labeled with ‘ns’ for non-significant), a significant difference between Sly-KD and WT samples was found with a P-value<0.05 (t-test). (c) Representation of SLY ChIP-Seq, its input, H3K4me3 ChIP-Seq, Kcr ChIP-Seq, H3K27me3 ChIP-Seq and H3K9me3 ChIP-seq. SLY co-localizes with active epigenetic marks H3K4me3 and Kcr at the TSS of H2afb3 (located in an intron of Pdch11x) but not of its autosomal-encoded homolog, H2afb1. (d) ChIP-qPCR experiments on WT and Sly-KD round spermatids using antibody against H2A.B3. The TSS of expressed genes was found enriched in H2A.B3 in Sly-KD compared to WT round spermatids (n=3 samples per genotype). The Y-axis represents the mean enrichment (%IP/input) normalized to the corresponding WT value. A significant increase in H2A.B3 level at the TSS was found in Sly-KD compared to WT samples when considering all tested genes (t-test, P<0.05)
