Figure 1.
Figure 1. Effect of manipulating STAT3 activities of mouse bone marrow cells on their hematopoietic repopulating ability. (A) Schematic drawings of the first 3 retroviral vectors used: MSCV-IRES-GFP (MIG, the control vector) and vectors encoding a dnStat3 cDNA or a Stat3-C cDNA. (B) Protein expression of STAT3-C or dnSTAT3 in transduced bone marrow cells analyzed 2 days after transduction. Arrows show the position of the proteins indicated. (C) Effect of altering the STAT3 activity of transplanted bone marrow cells on their in vivo repopulating ability. Bone marrow cells from 5-FU–pretreated Pep3b mice were transduced and then transplanted immediately into irradiated B6 recipients (1 to 2 × 105 cells plus 105 normal B6 bone marrow cells per recipient). The values shown are the mean ± SEM of the percent transduced (GFP+) cells detected in the peripheral-blood WBCs in 3 independent experiments (MIG, 16 mice; dnSTAT3, 11 mice; STAT3-C, 13 mice). (D) Lineage distribution of WBCs regenerated from transplanted Stat3-C–or dnStat3-transduced bone marrow cells analyzed 16 weeks after transplantation. GFP+ cells were analyzed for expression of myeloid (Mac1/Gr1) and B-lymphoid (B220) markers. The number of GFP+ T cells present was estimated as the number of B220– cells within the total lymphoid population (cells with low side and forward light-scattering properties). Values shown are the mean ± SEM of values from 5 individual mice in a representative experiment. (E) Multilineage reconstitution of mice by Stat3-C–transduced cells. Single-cell suspensions were prepared from the bone marrow (BM), thymus (Thy), spleen (Spl), and peripheral blood (PB) of mice that received a transplant of MIG- or Stat3-C–transduced bone marrow cells 36 weeks earlier, and the percentage of GFP+ cells was then determined. Values shown are the mean ± SEM (4 mice in each group). (F) Southern blots showing common proviral integrants in different tissues repopulated with Stat3-C–transduced cells. Genomic DNA was extracted, digested with Hindlll, and then probed with a GFP probe.

Effect of manipulating STAT3 activities of mouse bone marrow cells on their hematopoietic repopulating ability. (A) Schematic drawings of the first 3 retroviral vectors used: MSCV-IRES-GFP (MIG, the control vector) and vectors encoding a dnStat3 cDNA or a Stat3-C cDNA. (B) Protein expression of STAT3-C or dnSTAT3 in transduced bone marrow cells analyzed 2 days after transduction. Arrows show the position of the proteins indicated. (C) Effect of altering the STAT3 activity of transplanted bone marrow cells on their in vivo repopulating ability. Bone marrow cells from 5-FU–pretreated Pep3b mice were transduced and then transplanted immediately into irradiated B6 recipients (1 to 2 × 105 cells plus 105 normal B6 bone marrow cells per recipient). The values shown are the mean ± SEM of the percent transduced (GFP+) cells detected in the peripheral-blood WBCs in 3 independent experiments (MIG, 16 mice; dnSTAT3, 11 mice; STAT3-C, 13 mice). (D) Lineage distribution of WBCs regenerated from transplanted Stat3-C–or dnStat3-transduced bone marrow cells analyzed 16 weeks after transplantation. GFP+ cells were analyzed for expression of myeloid (Mac1/Gr1) and B-lymphoid (B220) markers. The number of GFP+ T cells present was estimated as the number of B220– cells within the total lymphoid population (cells with low side and forward light-scattering properties). Values shown are the mean ± SEM of values from 5 individual mice in a representative experiment. (E) Multilineage reconstitution of mice by Stat3-C–transduced cells. Single-cell suspensions were prepared from the bone marrow (BM), thymus (Thy), spleen (Spl), and peripheral blood (PB) of mice that received a transplant of MIG- or Stat3-C–transduced bone marrow cells 36 weeks earlier, and the percentage of GFP+ cells was then determined. Values shown are the mean ± SEM (4 mice in each group). (F) Southern blots showing common proviral integrants in different tissues repopulated with Stat3-C–transduced cells. Genomic DNA was extracted, digested with Hindlll, and then probed with a GFP probe.

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