Comparison of the transcriptomic effects of Foxo1-KO and treatment with AS1842856 in murine BCR::ABL1⁺ B-ALL. (A) RNA-sequencing was performed in BCR::ABL1⁺ Cre-ERT2 Foxo1^fl/fl murine B-ALL cells treated with either 200 nM 4-OHT or an equivalent amount of vehicle EtOH or with 70 nM AS1842856 or equivalent amounts of DMSO vehicle. RNA was isolated after 24 and 48 hours. Analysis was performed using the DESeq2 workflow in RStudio (version 2024.12.1). The unsupervised hierarchical clustering of all genes found in all samples is shown (supplemental File 1). (B) Hierarchical clustering of genes significantly (P < .05; log₂[FC] > 1.4) regulated was performed in both groups at 24 and 48 hours after respective treatment. (C) Volcano plot of differentially expressed genes. Significance threshold was set to adjusted P <.05 and log₂(FC) >1.5. Significantly downregulated genes are shown in blue; significantly upregulated genes are shown in red. Canonical FOXO1 targets are highlighted and labeled in black. (D) Differentially expressed genes from days 1 and 2 of either Foxo1-KO or AS1842856-treated samples were pooled and subjected to GSEA for “Hallmarks” gene sets using GSEA (version 4.3.3; supplemental File 2). (E) Differentially regulated genes in BCR::ABL1⁺ murine B-ALL after treatment with 70 nM AS1842856 or genetic Foxo1-KO were subjected to Venn analysis. Exclusive and shared genes were cross-referenced to the ChEA 2022 database using Enrichr (supplemental Materials and methods). ChEA, ChIP-X Enrichment Analysis; FC, fold change.