1. by Xiaowu Zhang, and Ruibao Ren
Bcr-Abl Efficiently Induces a Myeloproliferative Disease and Production of Excess Interleukin-3 and Granulocyte-Macrophage Colony-Stimulating Factor in Mice: A Novel Model for Chronic Myelogenous Leukemia
by Xiaowu Zhang, and Ruibao Ren
Blood
Volume 92(10):
November 15, 1998
©1998 by American Society of Hematology

2. The retrovirus construct used to transduce thebcr-abl/p210 and gfp genes.
The retrovirus construct used to transduce thebcr-abl/p210 and gfp genes. The construct MSCV-bcr-abl/p210-IRES-gfp was made as described in Material and Methods. LTR, long terminal repeat. Enzyme abbreviations: RI,EcoRI; N, Nco I; S, Sal I.
Xiaowu Zhang, and Ruibao Ren Blood 1998;92:
©1998 by American Society of Hematology

3. Survival of recipient mice after transplantation of bone marrow cells that were infected with retroviruses containing various genes as indicated.
Survival of recipient mice after transplantation of bone marrow cells that were infected with retroviruses containing various genes as indicated. The number of mice (n) used in the experiment for each retrovirus construct is indicated.
Xiaowu Zhang, and Ruibao Ren Blood 1998;92:
©1998 by American Society of Hematology

4. Pathological analysis of the Bcr-Abl–BMT mice.
Pathological analysis of the Bcr-Abl–BMT mice. (a) Peripheral blood smear from a primary Bcr-Abl–BMT mouse with the myeloproliferative disorder (original magnification × 600). The peripheral blood smear was stained with LeukoStat from Fisher. (B through D) Histological sections (original magnification × 400) of the spleen (b), liver (c), and lung (d) from a primary Bcr-Abl–BMT mice with the myeloproliferative disorder. Histological sections were stained with hematoxylin and eosin. Arrows in (c) point out mitotic hematopoietic cells.
Xiaowu Zhang, and Ruibao Ren Blood 1998;92:
©1998 by American Society of Hematology

5. Immunophenotyping of leukemic cells from Bcr-Abl–BMT mice by flow cytometry.
Immunophenotyping of leukemic cells from Bcr-Abl–BMT mice by flow cytometry. Two-parameter dot-plots show expression of lineage-specific antigens versus GFP as indicated. (Top panel) The expression of B220, Thy1.2, Ter119, Mac-1, and Gr-1 versus GFP, as indicated, in the peripheral WBCs from a primary Bcr-Abl–BMT mouse with the myeloproliferative disorder. (Bottom panel) The expression of Mac-1 versus GFP in the peripheral WBCs, spleen, liver, lung, and bone marrow from a secondary Bcr-Abl–BMT mouse with the myeloproliferative disorder.
Xiaowu Zhang, and Ruibao Ren Blood 1998;92:
©1998 by American Society of Hematology

6. Xiaowu Zhang, and Ruibao Ren Blood 1998;92:3829-3840
©1998 by American Society of Hematology

7. Xiaowu Zhang, and Ruibao Ren Blood 1998;92:3829-3840
©1998 by American Society of Hematology

8. Xiaowu Zhang, and Ruibao Ren Blood 1998;92:3829-3840
©1998 by American Society of Hematology

9. Xiaowu Zhang, and Ruibao Ren Blood 1998;92:3829-3840
©1998 by American Society of Hematology

10. Expression of Bcr-Abl/p210 protein in the peripheral WBC from a Bcr-Abl–BMT mouse with the myeloproliferative disorder and expression of the wild-type and kinase-negative mutant Bcr-Abl/p210 proteins in NIH 3T3 cells.
Expression of Bcr-Abl/p210 protein in the peripheral WBC from a Bcr-Abl–BMT mouse with the myeloproliferative disorder and expression of the wild-type and kinase-negative mutant Bcr-Abl/p210 proteins in NIH 3T3 cells. (A) Peripheral WBC from two Bcr-Abl–BMT mice with the myeloproliferative disorder (BMT4.18 and BMT5.8) were lysed directly in SDS sample buffer. The total cell lysates were subjected to Western blot analysis with anti-Abl monoclonal antibody Ab-3. The position of Bcr-abl/p210 and endogenous c-Abl is indicated. Total cell lysates from uninfected NIH3T3 and NIH3T3 infected with retrovirus containing the bcr-abl/p210 gene were used as negative and positive controls, respectively. (B) NIH 3T3 cells infected with equal amounts (1 mL) of MSCV-IRES-gfp (lane 1), MSCV-bcr-abl/p210-IRES-gfp (lane 2), and MSCV-K1176R-IRES-gfp (lane 3) retroviral supernatants of equal titer were subjected to Western blot analysis with anti-Abl monoclonal antibody Ab-3.
Xiaowu Zhang, and Ruibao Ren Blood 1998;92:
©1998 by American Society of Hematology

11. Detection of gfp in the genomic DNA of the peripheral WBCs from a Bcr-Abl–BMT mouse with the myeloproliferative disorder.
Detection of gfp in the genomic DNA of the peripheral WBCs from a Bcr-Abl–BMT mouse with the myeloproliferative disorder. Genomic DNA isolated from the unsorted WBCs (lane 1) and sorted GFP-negative (lane 2) and GFP-positive (lane 3) cells was subjected to PCR analysis using a mixture of primers that amplify the gfpgene (850 bp) and intron-3 of the mouse c-abl gene (500 bp). The amplified c-abl product was used as an internal positive control for the genomic DNA PCR.
Xiaowu Zhang, and Ruibao Ren Blood 1998;92:
©1998 by American Society of Hematology

12. Detection of gene expression of cytokines in bone marrow of transplanted mice by RT-PCR.
Detection of gene expression of cytokines in bone marrow of transplanted mice by RT-PCR. Bone marrow cells from Bcr-Abl–BMT mice (BMT5.1 [lanes 1 and 2], BMT5.4 [lanes 3 and 4], and BMT5.6 [lanes 5 and 6]) and a vector-BMT mouse (BMT5.35 [lanes 7 and 8]) was collected, and RNA was extracted as described in Materials and Methods. The RT-PCR products generated with (RT+) or without (RT−) reverse transcriptase were subjected to agarose gel electrophoresis, stained with ethidium bromide, and photographed by Gel Doc 1000 (Bio-Rad, Hercules, CA) with inverse gray scale. The RT-PCR products of IL-3, GM-CSF, and SCF are indicated. GAPDH was used as a control for the quality and quantity of the RNA samples and the RT-PCR reactions.
Xiaowu Zhang, and Ruibao Ren Blood 1998;92:
©1998 by American Society of Hematology

13. Detection of gene expression of IL-3 and GM-CSF in GFP-positive and GFP-negative bone marrow cells from Bcr-Abl–BMT mice.
Detection of gene expression of IL-3 and GM-CSF in GFP-positive and GFP-negative bone marrow cells from Bcr-Abl–BMT mice. Bone marrow cells from Bcr-Abl–BMT mice (BMT8.40 [lanes 1 through 4] and BMT8.41 [lanes 5 through 8]) were isolated and treated with ACK to lyse RBC. GFP-positive and GFP-negative bone marrow cells were sorted by flow cytometry and their RNA was extracted as described in Materials and Methods. The RT-PCR was performed and analyzed as in Fig8. The RT-PCR products of IL-3, GM-CSF, GFP, and GAPDH are indicated.
Xiaowu Zhang, and Ruibao Ren Blood 1998;92:
©1998 by American Society of Hematology