Figure 3
Figure 3. Effect of Baf250aE2E3 on HSC activity. (A) Homozygosity at Baf250aE2E3 allele enhances the competitive repopulation ability of mutant FL cells. WT, Baf250aE2E3, or Baf250aE2E3/E2E3 (CD45.2+) FL cells were mixed in a 1:4 ratio with CD45.1+ WT FL cells, and transplanted into lethally irradiated CD45.1+ recipient mice. Contribution of donor cells in peripheral blood of recipient mice was assessed at different times after transplantation. (B) BrdU incorporation profile of the CD150+CD48−Lin− cell population was assessed after 30 minutes in vivo BrdU pulse. Cell numbers are mean (± SD) values obtained from 3 developmentally matched FL. (C) Limiting dilution analysis for estimation of CRU frequency in WT and mutant FL (2 FL populations and 2 cohorts of recipient mice for each genotype). (D) Lympho-myeloid repopulation of recipient mice transplanted 20 weeks before with 100 000 WT or 10 000 Baf250aE2E3/E2E3 FL cells. Representative FACS profiles demonstrate comparable contributions of the transplant cell populations to repopulation of myeloid (Ly6G+), B lymphoid (CD45R+), and T lymphoid (CD4+) PBL. Average value in each quadrant represents number determined for mice (n = 5 in each group). (E) Proliferation potential of the transplanted HSCs. MAS, or proportion of PBL generated by individual transplant test CRU, was determined for groups of mice (n = 12) transplanted with low numbers (1-3) of Ly45.2+ WT or Baf250aE2E3/E2E3 FL CRU, together with 10 BM-derived competitor Ly45.1+CRU. (F) Contributions of the transplanted WT and Baf250aE2E3/E2E3 CRUs to regeneration of adult HSC pool. Proportions of test CD45.2+ and competitor CD45.1+-derived CD117+Ly6A/E+ (CD135−Lin− cells) in the BM of recipients described for Figure 1E were determined at 24 weeks after transplantation. Results represent mean (± SD) determined for each group of recipients (n = 5).

Effect of Baf250aE2E3 on HSC activity. (A) Homozygosity at Baf250aE2E3 allele enhances the competitive repopulation ability of mutant FL cells. WT, Baf250aE2E3, or Baf250aE2E3/E2E3 (CD45.2+) FL cells were mixed in a 1:4 ratio with CD45.1+ WT FL cells, and transplanted into lethally irradiated CD45.1+ recipient mice. Contribution of donor cells in peripheral blood of recipient mice was assessed at different times after transplantation. (B) BrdU incorporation profile of the CD150+CD48Lin cell population was assessed after 30 minutes in vivo BrdU pulse. Cell numbers are mean (± SD) values obtained from 3 developmentally matched FL. (C) Limiting dilution analysis for estimation of CRU frequency in WT and mutant FL (2 FL populations and 2 cohorts of recipient mice for each genotype). (D) Lympho-myeloid repopulation of recipient mice transplanted 20 weeks before with 100 000 WT or 10 000 Baf250aE2E3/E2E3 FL cells. Representative FACS profiles demonstrate comparable contributions of the transplant cell populations to repopulation of myeloid (Ly6G+), B lymphoid (CD45R+), and T lymphoid (CD4+) PBL. Average value in each quadrant represents number determined for mice (n = 5 in each group). (E) Proliferation potential of the transplanted HSCs. MAS, or proportion of PBL generated by individual transplant test CRU, was determined for groups of mice (n = 12) transplanted with low numbers (1-3) of Ly45.2+ WT or Baf250aE2E3/E2E3 FL CRU, together with 10 BM-derived competitor Ly45.1+CRU. (F) Contributions of the transplanted WT and Baf250aE2E3/E2E3 CRUs to regeneration of adult HSC pool. Proportions of test CD45.2+ and competitor CD45.1+-derived CD117+Ly6A/E+ (CD135Lin cells) in the BM of recipients described for Figure 1E were determined at 24 weeks after transplantation. Results represent mean (± SD) determined for each group of recipients (n = 5).

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