MOESM6 of DNA replication dynamics of vole genome and its epigenetic regulation

Additional file 7. HBO1 targeting to the heterochromatic block of the H3K27me3 decorated X chromosome leads to prolongation of its DNA replication duration. (A) Schematic representation of targeting assay to specifically target HBO1 to the heterochromatic block of the H3K27me3 enriched X chromosome. Cells were transiently transfected with GFP-tagged HBO1 and GBP-macroH2A1. Upon co-expression, both counterparts of the targeting assay strongly interact, leading to the targeting of HBO1. Targeting was validated by immunostaining with H3K27me3 antibody as a hallmark of facultative heterochromatin. DNA (blue), GFP-HBO1 (green), H3K27me3 (red) and merge as an overlay of all three channels. Scale bar 5 µm. (B) Site-directed targeting of HBO1 to the H3K27me3-enriched X chromosome leads to a significant prolongation of DNA replication duration of the X*. Specificity of the targeting approach was validated by the fact that there was no significant difference in H3K9me3 decorated X replication duration upon X* targeting.