Cdc73 shares ETS1 and Notch-driven pathways. (A) Rosa26CreER^T2Cdc73^f/f T-ALL cells (969 and 970) and control Rosa26CreER^T2 T-ALL cells derived from tumors in Figure 2C were treated with 3 nM OHT to delete Cdc73 and measured for growth. Fold expansion = day 3 or day 6 cell count / day 0 cell count. (B) Venn diagram showing 1062 Cdc73 target genes shared by both Cdc73^f/f T-ALL cell lines from panel A. Target genes defined as fold change (FC) > 1.5; Padj < .05 in Bru-seq counts at 30 hours after OHT addition in both Cdc73^f/f cell lines but not in controls. (C-D) GSEA using the top 258 ETS1-induced gene list¹⁷ of Cdc73-induced genes in 969 (C) and 970 (D) cells. (E-H) GSEA analyses showing the top 6 Hallmark pathways enriched for Cdc73- (E-F), ETS1- (G) and Notch- (H) induced target genes. ETS1 and Notch target genes were previously described in human THP-6 T-ALL cells.¹⁷ Pathways shared across the 4 analyses are highlighted in blue. (I-L) GSEA using the MSigDB C2 Kauffman_DNA_repair gene list of Cdc73-induced genes (I-J), ETS1-induced genes (K), and Notch-induced genes (L). (M-N) Venn diagram showing overlap of Cdc73-induced and ETS1-induced (M) or Notch-induced (N) genes in the Kauffman_DNA_repair gene list. (O-T) Volcano plots of significance vs Bru-seq (O-P,S) or RNA-seq (Q-R,T) Log2FC data showing the control vs OHT (Cdc73^Δ/Δ) comparison and highlighting important genes in nonhomologous end-joining (NHEJ; O,Q), homologous recombination (HR; P,R), and OXPHOS (S-T) pathways in 969 T-ALL cells (O-P,S) and AML cells (Q-R,T) on background of all genes (gray) giving average RPKM > 0.8. RNA-seq analysis of Control and Cdc73-deleted AML cells were obtained from.³⁵ ∗∗P < .01; ∗∗∗∗P < .0001. DMSO, dimethyl sulfoxide; EtOH, ethyl alcohol; GSI, gamma secretase inhibitor; RPKM, reads per kilobase per million.