HyperD-ALL cells are low proliferative and show a delay in early mitosis. (A) Heat map of the top 50 genes more differentially expressed between HyperD (n = 58) and non-HyperD (n = 30) B-ALL samples. (B-C) Top 20 statistically significant upregulated (B) or downregulated (C) biological pathways identified using GSEA for the genes differentially expressed in HyperD vs nonHyperD-ALL patients. Colored bars represent normalized enrichment scores (NES). P values are shown. (D) Sixteen-day proliferation curves for the indicated cell lines; n = 3 independent experiments. (E) SubG₀/SubG₁ apoptotic levels identified by FACS for the indicated cell lines; n = 3 independent experiments. (F) Cell cycle analysis for the indicated cell lines. Left, representative cell cycle FACS analysis. Right, frequency of cells in G₂/M analyzed; n = 3 independent experiments. (G) Representative DNA-Kinetochore-spindle IF staining (DNA, ACA, tubulin, and pericentrin) identifying the different mitotic phases in B-ALL cell lines. The SAC identifies the transition from early to late mitosis. Scale bar, 10 µm. (H-I) Mitosis progression in B-ALL cell lines. Progression from early to late mitosis (H), and frequency of cells at the indicated mitotic phases (I); n = 4 independent experiments. Graphs represent the mean, and error bars represent the standard error of the mean. *P < .05, **P < .01 (2-way analysis of variance [ANOVA]). A, anaphase; CK, cytokinesis; M, metaphase; P, prophase; PM, prometaphase; T, telophase.