Figure 2. Sequencing results of 5hmC in normal, abnormal, and globozoospermia sperm genomes.
(A) Dot blot detection of 5hmC in sperm DNA with positive and negative control. 400 ng, 200 ng, 100 ng and 50 ng genomic DNA purified from abnormal and globozoospermia sperm were blotted on Hybond-N+ nylon-based membrane and immune-blotted using anti-5-hydroxymethylytosine antibody. Synthesized oligonucleotides with and without 5hmC modification were included as positive and negative controls. (B) Mapping results of 5hmC sequencing in normal, abnormal, and globozoospermia sperm genomes. Raw reads were aligned to human UCSC hg19 and peaks calling using MACS (P < 10−5, fold enrichment > 10). (C) 5hmC peak numbers in normal, abnormal, and globozoospermia sperms. 20486, 38282 and 19354 peaks were identified in normal, abnormal, and globozoospermia sperm, respectively. (D) Average value of 5hmC fold enrichment in normal, abnormal, and globozoospermia sperm.