Figure 3.

Epigenetic Changes in the BU-1 Promoter Are Dependent on HU and the +3.5 G4 Motif

(A) H3K4me3 and H3K9me3 in untreated wild-type cells and cells treated with 150 μM for 48 hr and for 7 days. The enrichment for each mark is normalized to total H3 and then to untreated. The enrichment with non-specific IgG is shown as a control. Positive and negative controls for each mark are shown with ChIP at a constitutively active locus, GAS41, and a heterochromatinized locus, LYSC, respectively. Error bars show SD for three independent ChIPs.

(B) H3K4me3 and H3K9me3 in BU1^ΔG4 cells, either untreated or treated with 150 μM for 7 days.

(C) Enrichment of γH2Ax in wild-type and BU1^ΔG4 after treatment with HU for 7 days. γH2Ax signal is normalized to H3 and then to the level in untreated cells of each condition. Error bars show SD for four independent ChIPs for wild-type and five for BU1^ΔG4 cells.

(D) H3K4me3 in Bu-1a^low clones isolated after 7 days in 150 μM HU. Each point represents the results of a single ChIP from a single clone of either untreated (solid circles) or treated (open circles) cells. The central bar shows the mean and whiskers the SD for H3K4me3 in the five samples normalized to H3 and then to the mean of the untreated wild-type.

(E) H3K9me3 in the same clones as shown in (D). Normalization and error bars are as in (D).