Fig. 4

Bcl11a-deficient HSCs showed significant proliferative changes in the HSC compartment. a Distribution of Bcl11a ^−/− HSCs (purple) and cell cycle reconstructed-Bcl11a ^+/+ HSC clusters (C1–C5) in a PCA loading plot of the first two principal components. b Boxplots comparing the transcriptional activity of G0/G1 stage (C1 and C2) and S/G2/M stage (C3 and C4) HSCs in the Bcl11a ^+/+ and Bcl11a ^−/− datasets, estimated by the total number of read counts normalized by ERCC size factor per cell. c Violin plots of gene expression of selected cyclin genes, progenitor markers and cell cycle stage-associated genes in Bcl11a ^+/+ and Bcl11a ^−/− HSCs. The black dots represent the mean expression for each gene. d Heatmap showing expression correlations of selected transcription regulators in the HSC compartment. Correlation coefficient was calculated by Spearman correlation coefficient and clustering was performed by complete linkage. Gene correlation cluster II (blue) is magnified. e Enriched GO terms in gene correlation cluster I [red in (d)]. f Validation of cell cycle changes in Bcl11a ^−/− HSCs by 5-bromodeoxyuridine (BrdU) staining. The purple box marks the BrdU⁺ fraction in the HSC compartment of different genotypes. FSC-A: Forward scatter area. g The dose-dependent changes in BrdU⁺ cell number in the HSC compartment in different Bcl11a genotypes. HSCs were harvested and sorted from adult mouse bone marrow 5 days after tamoxifen-induced Bcl11a ablation. *p < 0.05, **p < 0.01, n = 3 mice for each group. Bcl11a ^+/−, CreERT2; Bcl11a ^+/flox (treated with tamoxifen); Bcl11a ^−/−, CreERT2; Bcl11a ^flox/flox (treated with tamoxifen). The error bar represented mean ± 1 standard deviation