CD43⁺ cell subsets: definition, kinetic profile, sorting and morphology. (A) The phenotype of CD43⁺ cells isolated on day 8 of H1/OP9 coculture. CD41a and CD235a were coexpressed (bottom left dot plot), and both in opposite manner to CD45 (top dot plots). Combination of CD41a-PE, CD235a-PE, and CD45-APC mAbs (bottom right dot plot) defines 3 major CD43⁺ subsets: (1) CD43⁺CD41a/CD235⁺CD45^–, (2) CD43⁺CD41a/CD235a^–CD45^–, and (3) CD43⁺CD41a/CD235a^– CD45⁺. Values within plots indicate percentage of cells in respective quadrants. Representative analysis is shown. (B) Kinetic analysis of indicated CD43⁺ subsets in H1/OP9 coculture represented as percentage of total CD43⁺ cells (left y1-axis). Dashed trend lines show parallel kinetics of indicated CFC types (right y2-axis). Results are the mean ± SD from 3 independent experiments. (C) Sorting strategy used for isolation of CD43⁺ subsets. A representative example of 7 independent experiments is shown (H1, n = 5; H9, n = 2). (D) Wright-stained cytospins of FACS-sorted CD43⁺ subsets (scale bar represents 5 μm). Images were captured with a Microphot-SA microscope (Nikon, Melville, NY) equipped with a 100×/1.40 oil-immersion objective lens, and were acquired through a DFC320 camera and Firecam 1.3 software (Leica Microsystems).