AMLs generated from CD34⁺ 38⁻ and GMP CB cells are transcriptionally similar to each other and to unmanipulated CB GMPs. (A) Unsupervised clustering (complete linkage) of transcript levels in GFP⁺ CD33⁺ (leukemic) cells isolated from NRG-3GS mice and compared with cell types purified by FACS from normal low-density CB cells. (B-C) REACTOME gene sets upregulated (B) and downregulated (C) in leukemic cells compared with normal CB GMPs. Reactome pathways enriched (P < .05 hypergeometric test) in differentially expressed genes were identified using ReactomePA, and networks clustering was performed based on the similarity of genes present in each pathway. (D-F) t-Distributed Stochastic Neighbor Embedding (t-SNE) analysis of the top 1000 variable genes of 4570 single GFP⁺CD33⁺ leukemic cells isolated from 2 AMLs generated from MYC-transduced CD34⁺38⁻ and 2 from MYC-transduced GMPs, following sample normalization and integration with Seurat v3, with t-SNE distributions of these cells shown by sample (D), by identified transcriptomic clusters (E), and cell cycle gene expression profiles (F).