Figure 3: ESL-1 controls HSPC proliferation by repressing TGFβ secretion.

(a) TGFβ levels in the BM of WT and Glg1^−/−-transplanted mice; n=8. (b) Micrographs of purified WT and Glg1^−/− LSK cells stained for pSMAD2/3 (red) and DAPI (blue). Scale bar, 5 μm. Data are from two independent experiments. (c) Expression of p18, p21 and p57 in sort-purified WT and Glg1^−/− LSK cells; n=6–8. (d) Representative contour plots of BrdU incorporation in WT and Glg1^−/− LSK cells and MP treated with anti-TGFβ or control antibody. Right, experimental design (top) and bar graphs (bottom) show the percentage of BrdU⁺ cells in the different groups; n=5. Each circle represents a mouse. (e) Representative plots and quantification of cell cycle analyses of WT and Glg1^−/− LSK cells and MP from mice treated with anti-TGFβ or control antibody; n=6. (f) Absolute numbers of the indicated progenitor populations in mice reconstituted with WT or Glg1^−/− marrow, and treated or not with anti-TGFβ antibody; n=5. (g) Recovery kinetics of leukocytes (WBC) and erythrocytes (RBC) in the blood of mice reconstituted with WT or Glg1^−/− BM after treatment with a single dose of 5-FU; n=10. *P<0.05; **P<0.01; ***P<0.001 determined by Student’s test (a,g), Wilcoxon matched-pairs (c) or one-way analysis of variance with Turkey’s test (d,e,f). Data are show as mean±s.e.m.