Figure 6.

Cell-Cycle Analysis and Functional Activity of G₀ LSKCD48−CD150+ Cells throughout the GFP Dilution Cascade

(A) Cell-cycle profiles of total LSKCD48−CD150+ cells or those at GFP 0, GFP 1, GFP 2, GFP 3, and GFP 4 by DNA content (DAPI) versus Ki67 staining. Cell-cycle phases were determined as G₀ Ki67^lowDAPI^2N, G₁ Ki67^hiDAPI^2N, S/G₂/M Ki67^hiDAPI ^{> 2N-4N}.

(B) Percentage of G₀, G₁, and S/G₂/M cells in total LSKCD48−CD150+ and those at GFP 0, GFP 1, GFP 2, GFP 3, and GFP 4 as determined by Ki67/DAPI staining (mean ± SD; n = 4 mice). Significance of G₀ cells determined by Student’s t test; ^∗p < 0.05; ^∗∗p < 0.01.

(C) Isolation of viable G₀ LSKCD48−CD150+ cells after 12 weeks chase for in vitro assays. LSKCD48−CD150+ cells were sorted first, followed by Py/Ho staining. G₀ cells (Py^lowHo^2N lower left quadrant) were isolated and separated into GFP 0, GFP 2, GFP 3, and GFP 4 subfractions.

(D) LTC-CAFC assay of G₀ LSKCD48−CD150+ cells at each GFP level.

(E) LTC-IC assay of G₀ LSKCD48−CD150+ cells at each GFP level. Significance of total colonies determined by Student’s t test; ^∗p < 0.05; mean ± SD; n = 3 separate cultures.