Figure 3: pTH2A is enriched in TSSs of sperm genome.
From: Identification of a variant-specific phosphorylation of TH2A during spermiogenesis
(a) Western blot analysis of sperm chromatin fractionated by MNase digestion. Antibodies used to detect each histone variant and modification are indicated. Number of biological replicates is shown as n. (b) An experimental procedure for ChIP-seq analysis using sperm nucleosome. (c) Enrichment of pTH2A replication 1 (Rep1), pTH2A replication 2 (Rep2), H3.3, H3.1/2, H3K4me3, and H3K27me3 in genomic elements. The mouse genome was divided into four elements: TSS region, 5 kbp around TSSs of all RefSeq genes; TES region, 5 kbp around transcriptional end sites (TES); genic region, region between TSS and TES; intergenic region, regions other than the former three elements. Data sets of H3.3, H3.1/2, H3K4me3, and H3K27me3 as well as their controls were obtained from Erkek et al.33. Enrichments (ChIP/input) are shown in log2. (d) Average profiles of each histone variant and modification enrichment ±5 kbp around TSS. H3K4me3 is excluded in the left panel to more clearly show other histone profiles. Ribbon: 95% confidence interval. (e) Heat map of ChIP-seq read density of each histone around TSSs. 2,500 genes were randomly selected and clustered into four groups: cluster1, 26.56% (664 genes); cluster2, 35.16% (879 genes); cluster3, 13.56% (339 genes); cluster4, 24.72% (618 genes). Genes are sorted by the average read density of pTH2A rep1 around TSSs (±1 kbp). Each row represents a gene centered on the TSSs. Relative read density denotes the scaled read density of each ChIP-seq data.
