Supplementary Figure S3 from Missense Mutations in Myc Box I Influence Nucleocytoplasmic Transport to Promote Leukemogenesis

Supplementary Figure 3. (A) Percentage peak overlap between replicate ChIP experiments for GFP+, mouse hematopoietic progenitor cells transduced with MycWT. (B) Distribution of peaks in GFP+ Linlow cells in relation to the nearest transcriptional start site (TSS) in GFP+ Linlow cells transduced with MycWT,MycP59Q and MycT58N vectors, EV (black line) relative to input controls (gray lines). (C) ChIP peaks motifs enrichment analysis indicating that both wild- type and mutant MYC proteins bind strongly to the E-box consensus CACGTG motif. (D) Gene Ontology analysis for MACS-called ChIP-seq peaks in GFP+ Linlow cells transduced with MycWT. (E) Heat map of absolute peaks read counts 5Kb upstream and downstream the TSSs for all expressed genes ranked by their signal in GFP+ Linlow cells transduced with EV, MycWT,MycP59Q and MycT58N vectors. The color scale is indicated at the bottom of each heatmap. (F) Western blot (top panel) and relative quantification (bottom panel) of cytoplasmic versus nuclear cellular lysates of GFP+ Linlow cells transduced with MycWT,MycP59Q and MycT58N vectors. The top bands represent MYC and the lower bands loading controls (ACTIN for cytoplasmic fraction and ORC2 for nuclear fraction). The bars represent average ± SD of MBI MYCMUT protein abundance in the nucleus and cytoplasmic fractions relative MYCWT (n = 3).