RNA-sequencing identifies cycling dysregulation in Cd53^−/− HSCs. (A) GO pathway analysis of DEGs (P_adj < .05) comparing mobilized splenic HSCs from G-CSF-treated Cd53^−/− and WT mice. (B) Heatmap of gene expression restricted to the known DREAM target genes in the indicated groups. (C-E) Volcano plots overlaying DREAM targets (yellow) over significantly downregulated (blue) and upregulated (red) genes between WT and Cd53^−/− HSCs from vehicle-treated BM (C), G-CSF-treated BM (D), and G-CSF-treated spleens (E). The point corresponding to Cd53 was excluded from panel E to better show the differential expression pattern; a complete volcano plot can be seen in supplemental Figure 2F. (F) Expression of p21, a known regulator of DREAM complex formation, is equivalently upregulated by G-CSF treatment in both WT and Cd53^−/− HSCs, as determined by bulk sequencing in panel A. (G) Overview of the DREAM complex regulation and function created using Biorender. Error bars represent mean ± SEM. ∗∗P < .01 by an unpaired Student t test.