Fig. 1: Glucocorticoid and B-cell pathways are co-expressed in healthy human B-cell progenitors.

A Flow cytometry gating strategy used to sort B-cell developmental populations (pre-pro-B: CD34⁺/CD38⁺/TdT⁺/CD24^-; pro-B: CD34^low/CD38⁺/TdT^-/CD24⁺; pre-B: CD34^-/CD38⁺/TdT^-/CD24⁺) in three healthy bone marrow donors for RNA-Seq analysis. B Canonical pathways are significantly enriched during development. Pathways highly correlated with BCR signaling were plotted; bold indicates BCR and glucocorticoid-related pathways. Heatmap is colored based on the z-score of IPA results. C Upstream regulators across development correlated with BCR complex; bold indicates BCR and glucocorticoid-related pathways. Heatmap is colored based on the z-score of IPA results. D Experimental workflow of healthy bone marrow donors analyzed by CyTOF. E Scaled median expression of B-cell related proteins and glucocorticoid receptor (GCR) in healthy pre-pro-B, pro-B, and pre-B cells. F GCR expression (Mean ± SEM) across cell cycle states (n = 3 healthy donors). Kruskal–Wallis nonparametric test is used to test significance (α = 0.05); G0 vs S p = 0.0285; G0 vs G2 p = 0.0358; S vs M p = 0.0225; G2 vs M p = 0.0255. G Mean percentage of each cell cycle phase in vehicle (ethanol) and dexamethasone (dex, 1 µM) treated cells. Paired t-test dexamethasone vs vehicle: G0: p = 0.0005, G1: p = 0.0021; S: p = 0.0789. H Percentage of live cells (cCASP3^-/cPARP^-) in vehicle (ethanol) and dexamethasone (1 μM) treated healthy cells (n = 3 healthy donors). Two-tailed paired t-test was used to test significance, p = 0.0742, not significant. Asterisks indicate p values as calculated by a two-tailed t-test (*p ≤ 0.5; **p ≤ 0.01; ***p ≤ 0.001; n.s. not significant). Source data are provided as a Source Data file. The experimental workflow was created with Biorender.com.