Figure 2

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.