PLCG1 is a target of AML1-ETO. (A) AML1 ChIP-sequencing analysis on normal CD34⁺ cells, (BCR-ABL⁺) K562, and (AML1-ETO⁺) SKNO-1 cells (https://genome.ucsc.edu). (B) Screenshot displaying changes in PLCG1 transcript levels based on RNA-sequencing (RNA-Seq, blue). Binding patterns of AML1-ETO (AE), AML1, JunD, CEBPa, LDB1, LMO2, PU.1, RNA-polymerase II (POLII), H3K27ac, and DHS at the PLCG1 locus in Kasumi-1 or patient-derived cells based on ChIP-sequencing and DNaseI-sequencing as well as conservation at the PLCG1 locus as aligned reads. Upper lines show promoter-Capture Hi-C (CHi-C) data generated in Kasumi-1 or patient-derived cells identifying DHSs interacting with the PLCG1 promoter. All data following inactivation of AML1-ETO (siAE; shAE) compared with nontargeting control (siMM; CTRL).^22-25 (C) Western blot analysis (left) and mRNA expression (right) in SKNO-1_Cas9-Blast cells (top) following CRISPR/Cas9 knockout using ETO-specific gRNA or a nontargeting control (sgLuc) and in Kasumi-1 cells (bottom) transduced with shRNA targeting AML1-ETO (AE) or empty vector control (shEV). n = 3 independent experiments; representative blot images are shown. (D) mRNA expression of PLCG1 (top) and AML1-ETO (bottom) in human embryonic stem (ES) cell-derived definitive hematopoietic progenitors expressing a Dox-inducible AML1-ETO fusion (data from 3 independent ES cell clones are shown). (E) Human ES-cell derived definitive hematopoietic progenitors expressing a Dox-inducible AML1-ETO fusion. ChIP-seq analysis displaying AML1-ETO binding at the PLCG1 locus (blue) without Dox (0 Dox) and after Dox treatment (5 ng/mL; 5 Dox) for 24 hours. Chromatin accessibility at the PLCG1 locus (green) after Dox treatment as indicated by ATAC-seq.