Dexamethasone treatment of B-ALL cells increases transcription of factors upstream of mTORC1. (A) NR3C1-KO NALM-6 cells overexpressing either WT NR3C1 or a mutant defective in DNA binding (NR3C1 R477H) were treated for 24 hours with either DMSO or 100 nM dexamethasone, and phospho-S6K levels were evaluated by western blot. NR3C WT and KO cells expressing an empty vector served as controls. (B) Cells were treated for 96 hours with either dexamethasone at various doses (0.32 nM, 1 nM, 3.2 nM, 10 nM, or 32 nM) or DMSO, and cell viability was assessed, as described in Figure 2D. The data presented are representative of multiple independent experiments. (C) NALM cells were treated for 6 hours with either DMSO or 100 nM dexamethasone and harvested for RNA-seq. Fold-changes (x-axis) in gene expression upon dexamethasone treatment for each gene and their respective P values (y-axis) are displayed in volcano plots. Genes with significantly changed expression after dexamethasone treatment are depicted in dark gray, and, among them, those relevant to the PI3K-AKT-mTORC1 pathway are shown in red. (D) Gene set enrichment analysis (GSEA) analysis demonstrating enrichment of a PI3K-AKT-mTORC1 pathway signature in dexamethasone-treated cells. (E) Dot plots demonstrate a correlation between RNA-seq and CRISPR screen results.