Figure 3
Figure 3. Greater short-term and long-term repopulating activities of Ptpn11D61G/+ BM cells. (A-B) BM cells (1 × 105) freshly harvested from 3-month-old Ptpn11D61G/+ and WT littermates were injected into sub-lethally irradiated C57BL/6 mice (n = 9 per group). Twelve days after the transplantation, colonies on the spleen (CFU-S) were counted. Spleens were photographed using a Canon PowerShot SD600 digital camera. (C) BM cells (test cells; 1 × 106) harvested from WT or Ptpn11D61G/+ littermates (CD45.2+) were transplanted with the same number of BoyJ (CD45.1+) BM cells (competitor cells) into lethally irradiated BoyJ (CD45.1+) mice (n = 5 per group). Test cell reconstitution (CD45.2+) was determined at 4, 8, 12, and 16 weeks by FACS analyses of peripheral blood cells of the recipient mice. (D) Sixteen weeks after transplantation, percentages of test cell–derived CD45.2+ cells in the whole BM population were determined (n = 5 per group). (E) Contributions of test cells (CD45.2+) to each lineage (Mac-1, Gr-1, B220, Ter119, and CD4 positive cells) in the BM were also quantified (n = 5 per group). (F) BM cells (2 × 106) harvested from the primary recipients were transplanted into lethally irradiated BoyJ (CD45.1+) mice. Sixteen weeks after the transplantation, BM cells of the secondary recipients were analyzed for the contribution of test cells in the BM (n = 5 per group). (G) Reconstitution from test cells in each lineage of the BM was determined as described in panel E (n = 5 per group). (H-I) BM cells harvested from WT or Ptpn11D61G/+ littermates were transplanted with the same number of BoyJ BM cells into lethally irradiated BoyJ mice (n = 5 per group) as in panel C. Sixteen weeks after the transplantation, recipient mice were killed and frequencies of LSK cells renewed from test cells (CD45.2+) in the BM (H) and spleen (I) were quantified by multiparameter FACS analyses as described in Figure 1. BM cells (2 × 106) harvested from primary recipient mice were transplanted into lethally irradiated BoyJ mice. Sixteen weeks after transplantation, frequencies of LSK cells renewed from test cells (CD45.2+) were quantified in the BM (J) and spleen (K) of the secondary recipient mice.

Greater short-term and long-term repopulating activities of Ptpn11D61G/+ BM cells. (A-B) BM cells (1 × 105) freshly harvested from 3-month-old Ptpn11D61G/+ and WT littermates were injected into sub-lethally irradiated C57BL/6 mice (n = 9 per group). Twelve days after the transplantation, colonies on the spleen (CFU-S) were counted. Spleens were photographed using a Canon PowerShot SD600 digital camera. (C) BM cells (test cells; 1 × 106) harvested from WT or Ptpn11D61G/+ littermates (CD45.2+) were transplanted with the same number of BoyJ (CD45.1+) BM cells (competitor cells) into lethally irradiated BoyJ (CD45.1+) mice (n = 5 per group). Test cell reconstitution (CD45.2+) was determined at 4, 8, 12, and 16 weeks by FACS analyses of peripheral blood cells of the recipient mice. (D) Sixteen weeks after transplantation, percentages of test cell–derived CD45.2+ cells in the whole BM population were determined (n = 5 per group). (E) Contributions of test cells (CD45.2+) to each lineage (Mac-1, Gr-1, B220, Ter119, and CD4 positive cells) in the BM were also quantified (n = 5 per group). (F) BM cells (2 × 106) harvested from the primary recipients were transplanted into lethally irradiated BoyJ (CD45.1+) mice. Sixteen weeks after the transplantation, BM cells of the secondary recipients were analyzed for the contribution of test cells in the BM (n = 5 per group). (G) Reconstitution from test cells in each lineage of the BM was determined as described in panel E (n = 5 per group). (H-I) BM cells harvested from WT or Ptpn11D61G/+ littermates were transplanted with the same number of BoyJ BM cells into lethally irradiated BoyJ mice (n = 5 per group) as in panel C. Sixteen weeks after the transplantation, recipient mice were killed and frequencies of LSK cells renewed from test cells (CD45.2+) in the BM (H) and spleen (I) were quantified by multiparameter FACS analyses as described in Figure 1. BM cells (2 × 106) harvested from primary recipient mice were transplanted into lethally irradiated BoyJ mice. Sixteen weeks after transplantation, frequencies of LSK cells renewed from test cells (CD45.2+) were quantified in the BM (J) and spleen (K) of the secondary recipient mice.

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