10.1371/journal.pone.0000306.g001 Chozhavendan Rathinam Chozhavendan Rathinam Christoph Klein Christoph Klein Developmental dichotomy of CLP1 and presumptive CLP2 progenitor cells evidenced by analysis of Gfi1<sup>−/−</sup> and Gfi1<sup>GFP/+</sup> mice. Public Library of Science 2007 dichotomy clp1 presumptive clp2 progenitor cells evidenced 2007-03-21 00:33:22 Figure https://plos.figshare.com/articles/figure/_Developmental_dichotomy_of_CLP1_and_presumptive_CLP2_progenitor_cells_evidenced_by_analysis_of_Gfi1_8722_8722_and_Gfi1_GFP_mice_/622002 <p>A) FACS plots indicating relative decrease of CLP1 (upper panels) and normal numbers of presumptive CLP2 (middle panels) in Gfi1<sup>−/−</sup> bone marrow cells. Cells were pregated on Lin<sup>−</sup>IL7Rα<sup>+</sup> cells (upper panels) and Lin<sup>−</sup>IL7Rα<sup>+</sup>Sca1<sup>+</sup>CD19<sup>−</sup> cells (middle panels), respectively. Bar diagrams indicating absolute numbers of CLP1 and CLP2 cells of Gfi1<sup>−/−</sup> and Gfi1<sup>+/+</sup> mice (lower panel, n = 3 mice). Lineage markers included CD4, CD8, CD11c, CD11b and B220. Flow-cytometric analysis was performed on pooled BM cells from 5 mice. Shown is a representative experiment of 3. B) FACS plots indicating relative increase of ETP in Gfi1<sup>−/−</sup> thymus. Lin<sup>−</sup>CD25<sup>−</sup>CD44<sup>+</sup> cells (upper panels) were gated (G1) and analysed for expression of c-kit and IL7Rα (lower panels). Experiments were performed on pooled thymi (n = 5 mice). Bar diagrams indicating absolute numbers of ETP of Gfi1<sup>−/−</sup> and Gfi1<sup>+/+</sup> mice (lower panel, n = 3 mice). The total number of thymocytes was decreased by ∼6 fold in Gfi1<sup>−/−</sup> mice. Shown is a representative experiment of 3. C) Transcriptional activity of Gfi1 locus in hematopoietic progenitor cells and thymic B cells. Pooled bone marrow cells or thymocytes (n = 5) were analyzed for GFP expression in the following populations: LSK (HSC), Lin<sup>−</sup>IL7Rα<sup>+</sup>Sca1<sup>low</sup>c-kit<sup>low</sup> (CLP1), Lin<sup>−</sup>IL7Rα<sup>+</sup>Sca1<sup>+</sup>c-kit<sup>−</sup>B220<sup>+</sup>CD19<sup>−</sup> (CLP2) and Lin<sup>−</sup>CD25<sup>−</sup>IL7Rα<sup>−</sup>CD44<sup>+</sup>c-kit<sup>+</sup> (ETP). Shaded histograms represent GFP fluorescence in Gfi1<sup>+/GFP</sup> cells, open histograms represent autofluorescence of Gfi1<sup>+/+</sup> cells. The GMFI (Geo Mean Fluorescence Intensity) of Gfi1<sup>+/+</sup> (above) and Gfi1<sup>+/GFP</sup> (below) cells is indicated. Data are representative of 3 independent experiments.</p>