Figure 3
From: Lineage-specific distribution of high levels of genomic
The dynamics of 5-hydroxymethylcytosine (5-hmC) in mouse post-implantational embryonic development. (A) Sagital sections of 10.5 days post coitum (dpc), 12.5 dpc and 15.5 dpc mouse embryos stained with Diagenode antibody to 5-hmC. Immunostaining for single-stranded DNA is shown in red. (B) Fragments of brain and tongue (indicated) embryonic sagital sections at 10.5 dpc, 12.5 dpc and 15.5 dpc stages (indicated). Corresponding slides were immunostained for 5-hmC in parallel with identical conditions and imaged with the same exposure times. Cell populations of embryonic tongue enriched in 5-hmC at 15.5 dpc are arrowed. (C) A schematic showing of the results of quantification of 5-hmC signal in embryonic brain and tongue at 10.5 dpc, 12.5 dpc and 15.5 dpc. Corresponding slides were immunostained for 5-hmC in parallel and imaged with the same exposure times. Mean values for mean signal intensities of 10 random measurements of parts of images shown in B are presented. The same central part of developing brain was quantified for all the stages analysed. Error is expressed as s.e.m. (D) The results of 5-hmC immunostaining of 3 adjacent sections of 12.5 dpc embryo brain region using primary antibody mix pre-incubated with PCR-produced DNA fragments where all the cytosines were replaced with either 5-hmC (5-hmC DNA) or 5-mC (5-mC DNA). Control staining without pre-incubation with any DNA is shown (no DNA). The 5-hmC immunostaining is specifically competed by 5-hmC- but not by 5-mC-containing DNA. Genomic DNA was visualised using single-stranded DNA antibody (indicated) used at a high titre. The experiments were performed using the Diagenode antibody.
