cbCD34 models recapitulate the phenotypes of NUP98r leukemia. (A) Experimental schema using cbCD34 models. (B) Colony-forming unit assays of cbCD34 models with empty control vectors or NUP98::NSD1, NUP98::KDM5A, or NUP98::HOXA9-expressing vectors. (C) Cell growth assays of cbCD34 models in liquid culture. (D) Flow cytometric analysis of cbCD34 models in liquid culture (top, CD34⁺; middle, CD11b⁺; bottom, CD41a⁺; population ratio, % in mCherry⁺ live cells). (E) Principal component analysis of RNAseq data from liquid culture. Colors indicate NUP98 fusions, and shapes indicate days after transduction. (F) Heat map showing expression of representative genes related to stemness or differentiation of hematopoietic cells. The colors of cells indicate expression levels normalized among samples, and genes are annotated on the left. (G) Comparison of differentially expressed genes in each cbCD34 model compared with empty vector controls at day 42. Venn diagram showing overlaps of highly expressed genes in each model (middle), and gene ontology term analyses of shared or specific differentially expressed genes are shown (left, right). Gray lines show −log10 FDR = 0.05. Data were obtained from 3 biological replicates (different lots of cord blood). In panels B-D, statistical tests were performed by a generalized linear mixed effect model with Poisson (B) and Gaussian (C-D) distributions, followed by comparison with empty vector control and the Benjamini-Hochberg adjustment, asterisks indicating adjusted P values, ∗P < .05. Error bars indicate mean ± standard error of the mean. FDR, false-discovery rate; PDX, patient-derived xenograft.