FIG 7.

Setd8 knockdown results in significant changes in chromatin architecture in the Gata2 locus. ChIP assays were performed in self-renewing Setd8 knockdown and scramble control ESREs. Data are presented as fold enrichment relative to total input control and represent the means and SEM from a minimum of three independent experiments. (A) Setd8 occupies the Gata2 1S promoter and +9.5 enhancer in scramble control ESREs. Setd8 knockdown is associated with loss of Setd8 occupancy in the Gata2 locus. The positive control is the beta actin intron, and the negative control is a region upstream of the Gata2 locus with no known function. (B) There is occupancy of H4K20me1 at the Gata2 1S promoter that is lost with Setd8 knockdown. The positive control is beta actin intron, and the negative control is a region upstream of the Gata2 locus with no known function. (C) Setd8 knockdown results in increased histone H4 acetylation at the Gata2 1S promoter. The positive control is the beta actin promoter. The negative control is the necdin gene, which is not expressed in erythroid cells.