Cell cycling and differentiation of HSCs are normal in arrhythmic Bmal1 deficient mice. A, B) Normal frequency of HSCs in the BM of 8-10-week-old Bmal1
−/− mice. CD34−KSL fractions were assessed by flow cytometry. A) Data shown are representative of CD34−KSL cells at ZT5 and ZT17. B) The mean percentages ± SDs of CD34−KSL cells at ZT5 (n = 4) and ZT17 (n = 3) of two independent experiments. C) Comparable frequency of quiescent cells in HSC populations. HSCs of Bmal1
+/+ and Bmal1
−/− mice were stained with Pyronin Y and analyzed by flow cytometry to give the mean percentages ± SDs of Pyronin Y− cells in the CD34−KSL populations at ZT5 and ZT17 (n = 3) of two independent experiments. D) Normal EdU incorporation in Bmal1
−/− CD34−KSL cells. EdU was administered orally to mice for 3 weeks, and EdU incorporation into HSCs was evaluated using a Click-iT EdU PB Flow Cytometry Assay Kit. Data shown are the mean percentages ± SDs of EdU+ cells in HSC populations (Bmal1
−/− mice; n = 6, Bmal1
−/− mice; n = 3). E) White blood cell differentiation in young (10-week-old) and aged (40-week-old) mice. Each stack in the bar represents a cell type percentage. Gr-1+, granulocytes; Mac-1+, macrophages; B220+, B cells; CD4+, CD4+ T cells; and CD8+, CD8+ T cells (n = 6) of four independent experiments.