Fig. 3: Cyclin D1 association with histones is governed by phosphorylation (H2B^S14).

A Representative example of histone array consisting of 384 unique histone modification combinations in duplicate. The array includes up to four different modifications on the same 19mer peptide. For the individual modifications, please see Supplemental Table 3. The Y axis is labeled alphabetically for a given histone in each row, with modifications labeled numerically on the X axis. As each spot is arrayed in duplicate, representative examples of cyclin D1 binding to modified histones as tested by histone-modification arrays and analyzed by active Motif software are shown. B Binding is representative of the average positive intensity over the average negative intensity of each modification. The correlation between densitometric analysis of duplicate interactions for each interaction from two separate arrays is shown as intensity left vs. intensity right. The intensity is highly reproducible between duplicate binding assessments (R² = 0.927). (P refers to the designation of the column on the array (H2B), and the number refers to the numerical labeling of the row). C Representative examples of histone arrays (384 unique histone modifications on the same 19mer peptide), with binding shown to GST-cyclin D1 (GST-CD1^WT), GST-CD1^ΔE, GST-KDM4A or GST-ctrl fusion proteins. D Representative and (E) quantitated binding, representing average positive intensity over the average negative intensity of each modification for H2B 19mer peptide. F Graphical quantification of cyclin D1 binding to post-translationally modified H2B^S14 19mer peptide.