Developmental kinetics and sorting strategy of CD34⁺ subsets. (A) Kinetic analysis of CD34⁺ subsets in H1/OP9 coculture. Indicated CD34⁺ subsets were analyzed by FACS within gated CD34⁺ population and are represented on the top graph as percentage of CD34⁺ cells (left y1-axis). Total CD34⁺ cells (%) are depicted by dashed trend line (right y2-axis). The bottom graph shows absolute numbers of respective CD34⁺ subsets. Results are the mean ± SD from 4 independent experiments. (B) Flow diagram of multiparameter MACS separation of CD34⁺ subsets showing a FACS analysis of target cell populations throughout the sorting procedure (see “Materials and methods” for details). Positive and negative MACS fractions are indicated by arrows marked with + and – circles, respectively. Values within dot plots indicate percentage of cells in respective quadrants. The representative experiment is shown.