Multilineage reconstitution after transplantation of CD41-GFP^lo cells into irradiated primary and secondary recipients. (A) The top panel analyzes donor-specific signals by PCR in the genomic DNA of unfractionated peripheral blood cells 1-3 months after transplantation and flow-sorted WKM cells 3 months after transplantation. The bottom panel depicts a plot of the forward and side scatter of WKM cells and the location, as determined by microscopic examination, of the major blood cell lineages. The pink ovals mark the windows for erythroid, myeloid, lymphoid, and various precursor populations in zebrafish WKM determined by examining Wright-Giemsa stains of cytospins derived from the flow-sorted cells. The percentages vary slightly with each WKM preparation but the ones shown here are representative. FSC-H indicates forward scatter; and SSC-H, side scatter. The bar chart in panel B shows the percentage of total genomic DNA extracted from flow-sorted cells that contains the donor-specific signal (GFP) gated according to the windows depicted in panel A. Two fish per group were examined 6 and 10 months after primary transplantation and 6 months after secondary transplantation. As shown, the donor contribution percentage varied among lineages, with time after transplantation and among individual fish.