1. Volume 3, Issue 4, Pages 364-366 (October 2008)
Mobilized Hematopoietic Stem Cell Yield Depends on Species-Specific Circadian Timing 
Daniel Lucas, Michela Battista, Patricia A. Shi, Luis Isola, Paul S. Frenette 
Cell Stem Cell 
Volume 3, Issue 4, Pages (October 2008)
DOI: /j.stem
Copyright © 2008 Elsevier Inc. Terms and Conditions

2. Figure 1
Circadian HSC/Progenitor Fluctuations in Mice Modulate G-CSF and AMD3100 Mobilization
(A) Representative FACS plot showing the frequency of LSK cells in G-CSF-mobilized murine peripheral blood at ZT5 or ZT13. Live cells were gated, and the Lineage− fraction was analyzed to detect the fraction of Sca-1+ and c-kit+ cells.
(B) Number of LSK cells per ml of blood at ZT5 (white bar) or ZT13 (gray bar) in mice mobilized with G-CSF. The black bars in (B)–(E) depict the number of LSK cells or CFU-C at ZT5 or ZT13 in the circulation of steady-state mice (Mendez-Ferrer et al., 2008). ∗p < 0.05, n = 5 mice. Mice received G-CSF (125 μg/kg) s.c. every 12 hr for 4 days; the last dose of G-CSF was administered 3 hr before blood collection.
(C) Number of CFU-C per ml of blood at ZT5 (white bar) or ZT13 (gray bar) in mice mobilized with G-CSF using the same protocol as in (B). ∗∗p < 0.01; n = 10 mice.
(D) Number of LSK cells per ml of blood at ZT5 (white bar) or ZT13 (gray bar) in mice mobilized with AMD3100. AMD3100 (5 mg/kg) was injected i.c. 1 hr before blood collection. ∗p < 0.05; n = 6–8 mice.
(E) Number of CFU-C per ml of blood at ZT5 (white bar) or ZT13 (gray bar) in mice mobilized with AMD3100. AMD3100 mobilization was performed as in (D). ∗p < 0.05; n = 10 mice.
(F) Percentage of CXCR4 expression signal relative to ZT13 (measured by flow cytometry) at ZT5 and ZT13 in bone marrow LSK cells from nonmobilized mice. n = 3; ∗p < 0.05.
(G) CXCR4 expression (normalized to Bmal-1+/+,+/− circadian time [CT] levels) in bone marrow LSK cells collected from Bmal-1+/+,+/− (CT5, n = 7; CT13, n = 6) or Bmal-1−/− mice (CT5, n = 3; CT13, n = 4) housed for 1 week in a 12 hr dark: 12 hr dark regime. CXCR4 oscillations were conserved in Bmal-1+/+,+/− mice (∗∗p < 0.01) in constant darkness but ablated in Bmal-1−/− mice.
(H) CFU-C per ml of blood at ZT5 in mice housed in a 12 hr light: 12 hr dark regime (LD) or after a 12 hr jet lag. To induce the jet lag, the light cycle was advanced 12 hr at ZT12 (17 hr before blood collection at ZT5). AMD3100 mobilization was performed as in (D). ∗∗p < 0.01; n = 5 mice per group.
(I) CFU-C per ml of blood at CT5 or CT13 in mice housed in a 12 hr light: 12 hr light regime for 3 weeks and mobilized with G-CSF as in (B). n = 5 mice per group. All data are represented as mean ± SEM.
Cell Stem Cell 2008 3, DOI: ( /j.stem )
Copyright © 2008 Elsevier Inc. Terms and Conditions

3. Figure 2
Circadian Fluctuations of Human Progenitors in Peripheral Blood
(A) Representative peripheral blood FACS analysis from healthy subjects to detect CD34+ CD38− cells in peripheral blood collected at 8:00 a.m. and 8:00 p.m. Cells were first gated on the CD34+ populations and then analyzed for the presence of CD38− cells. Average number of CD34+ cells (B) and CD34+CD38− cells (C) per ml of blood at indicated times; n = 9; ∗∗∗p < (D) Total colony-forming units in culture (CFU-C) detected in the peripheral blood from same donors; ∗∗∗p < (E) Number of CD34+ cells recovered per blood volume processed (l) per weight (kg) of healthy donors that were mobilized using G-CSF (10 μg/kg every morning for 5 days) for allogeneic bone marrow transplantation at Mount Sinai Medical Center between 2001 and Subjects were grouped according to the apheresis half-time. ∗∗∗p <
Cell Stem Cell 2008 3, DOI: ( /j.stem )
Copyright © 2008 Elsevier Inc. Terms and Conditions