Hematopoietic specification of KMT2A-rearranged iPSCs induces a transcriptionally distinct profile. (A) Schematic of the generation of HSPCs from AML- and control-iPSC cell lines. (B) Representative images of hematopoietic cultures of control- and AML-iPSCs depicting the generation of HSPCs following 13 days of differentiation. (C) Representative flow cytometry diagrams of hematopoietic cell populations from control- and AML-HSPCs after 13 days of differentiation, and (D) quantification of the viability and expression of key hematopoietic populations by flow cytometry (n = 3). Data are presented as mean ± standard error of the mean (SEM) of percent cells in hematopoietic cultures and analyzed using unpaired t test with Holm-Šídák multiple comparisons test. (E) Principal component analysis plot from RNA-seq of iPSCs (triangles) and day 13 HSPCs (circles) from AML and control cell lines, showing the first 2 principal components (n = 3). (F) Scatterplot of differential expression during hematopoietic differentiation, comparing log₂ fold changes (HSPCs vs iPSCs) in control (x-axis) and AML (y-axis). Genes were classified into categories including activated in both AML and control (green), activated in AML only (orange), actively repressed in AML (red), failed to activate in AML (blue), downregulated in both AML and control (purple), and ns/other (gray). ∗P ≤ .05. ns, not significant.