Single-cell transcriptome analysis of JMJD1C Jumonji domain–mutated MLL-AF9 leukemia cells. (A-B) T-distributed stochastic neighbor embedding plot of single-cell gene-expression data of mouse MLL-AF9-Cas9 leukemia cells 7 days after transduction with sgRNA against Renilla or JMJD1C JmjC domain (Jumonji). (A) Cell phenotype. (B) Cell clusters identified within phenotypes. The numbers of cells are indicated in parenthesis. (C) Cyclone cell cycle status. (D) Expression level of Somerville LSC³⁴  signature. (E) Monocle single-cell trajectory analysis. (F) SCENIC regulon analysis. Black bars indicate that a regulon is activated within a cell, across all clusters identified in panel B. (G-H) Decomposed Z score of MAST analysis on C6 oncogenic signature from the Molecular Signature Database (all Renilla cells vs all Jumonji cells). (G) Pink: enrichment in Jumonji; green: enrichment in Renilla sample, and hematopoietic fingerprint gene sets (Jumonji 2 cells vs Jumonji 3 cells). (H) Pink: enrichment in Jumonji 2; green: enrichment in Jumonji 3. GSVA analysis of top KRAS pathway (I; identified in panel G) and RAS/MAPK and JAK-STAT pathways (J). P < .01 for all pairwise comparisons in panel D, with the exception of Renilla 1 vs Renilla 2 (not significant), unpaired 2-tailed Student t test with Bonferroni correction. Gran, granulocytes; Mye, myeloid; Mono, monocytes; NK, natural killer cells.