Fig. 3.

Myb is required for distal enhancer activation and Gata1 binding in EryD. (A and B) Comparison of EryP-/EryD-specific Tal1 peaks and their genomic distribution (A). Corresponding association study of EryP-/EryD-specific Tal1 peaks and gene expression is shown in B. (C–E) Comparison of EryP-/EryD-specific Gata1 peaks and their genomic distribution (C). Corresponding association studies of EryP-/EryD-specific Gata1 peaks and gene expression are shown at genome-wide (D), proximal (E, Upper) and distal (E, Lower) regions. (F) Scatter plot of the difference of motif enrichment scores (y axis) and gene expression (x axis) reveals Myb (red spot) as an EryD-specific transcription factor. y axis represents the log₂ fold change of the percentage of EryP-/EryD-specific enhancers with motifs, while x axis represents the log₂ fold change of gene expression of the cognate TFs. Broken lines indicate threshold at fold change of 1.5 on motif enrichment (y axis) and threshold at fold change of 4 of gene expression (x axis). (G) Heatmaps of the normalized ChIP-seq reads of Gata1, H3K27ac ChIP-seq, p300, and Myb, centered around the Gata1 peak summits in EryP and EryD. EryP-/EryD-specific Gata1 peaks were separated into proximal (purple) and distal (green) regions. Myb and p300 ChIP-seq were performed in E13.5 fetal liver. (H and I) ChIP-seq signal density plots of Gata1 (H) and H3K27ac (I) in MEL cells treated with control shRNA (Ctrl shRNA) or Myb shRNA. Signals included in plots were restricted to EryD-specific distal regions as shown in Fig. 3G. y axis is normalized ChIP-seq reads, log₂(RPKM + 1).