Figure 2
Figure 2. HIF1α is essential for CML development. (A) Deletion of HIF1α in hematopoietic cells. Mice carrying the floxed HIF1α allele were crossed with Vav-Cre transgenic mice in which the Vav promoter drives the expression of the Cre recombinase. RT-PCR analysis showed that HIF1α was undetectable in sorted HSCs (Lin−Sca-1+c-Kit+) from HIF1αflox/flox-Vav-Cre mice. (B) Kaplan-Meier survival curves for primary and secondary recipients of empty vector or BCR-ABL–transduced BM cells from WT or HIF1α−/− donor mice. For secondary BM transplantation, BM cells from primary control and CML recipient mice which received empty vector or BCR-ABL–transduced WT and HIF1α−/− BM cells were analyzed by FACS, and BM cells containing equal number of WT or HIF1α−/− GFP+Lin−Sca-1+c-Kit+ cells along with 2 × 105 WT BM cells (CD45.1) were transplanted into each lethally irradiated secondary recipient mouse. (C) FACS analysis of GFP+Gr-1+ cells in PB of primary and secondary recipients of empty vector or BCR-ABL–transduced BM cells from WT or HIF1α−/− donor mice (n = 5). Mean values (± SD) are shown. (D) The total numbers of GFP+Gr-1+ cells in PB of primary and secondary recipients of empty vector or BCR-ABL–transduced BM cells from WT or HIF1α−/− donor mice (n = 5). Mean values (± SD) are shown. *P < .05; **P < .01; ***P < .001. (E) The spleen weight of primary and secondary recipients of empty vector or BCR-ABL–transduced BM cells from WT or HIF1α−/− donor mice (n = 12). Mean values (± SD) are shown; ***P < .001. (F) Gross appearance of the lungs and spleens showed severe lung hemorrhages and splenomegaly in secondary recipients of WT LSCs but not HIF1α−/− LSCs at day 14 after BMT, compared with those from recipients of WT and HIF1α−/− HSCs. (G) H&E staining of tissue sections from lung and spleen of secondary recipients. The scale bar represents 50 μm. (H) Kaplan-Meier survival curves for the secondary BM transplantation. BM cells from WT or HIF1αflox/flox mice were transduced with BCR-ABL-iCre-GFP retrovirus, and transplanted into lethally irradiated recipient mice. CML cells without HIF1α failed to induce CML disease in the secondary BMT. (I) The percentages of leukemia cells in PB at day 14 in WT and HIF1α−/− secondary CML mice.

HIF1α is essential for CML development. (A) Deletion of HIF1α in hematopoietic cells. Mice carrying the floxed HIF1α allele were crossed with Vav-Cre transgenic mice in which the Vav promoter drives the expression of the Cre recombinase. RT-PCR analysis showed that HIF1α was undetectable in sorted HSCs (Lin−Sca-1+c-Kit+) from HIF1αflox/flox-Vav-Cre mice. (B) Kaplan-Meier survival curves for primary and secondary recipients of empty vector or BCR-ABL–transduced BM cells from WT or HIF1α−/− donor mice. For secondary BM transplantation, BM cells from primary control and CML recipient mice which received empty vector or BCR-ABL–transduced WT and HIF1α−/− BM cells were analyzed by FACS, and BM cells containing equal number of WT or HIF1α−/− GFP+Lin−Sca-1+c-Kit+ cells along with 2 × 105 WT BM cells (CD45.1) were transplanted into each lethally irradiated secondary recipient mouse. (C) FACS analysis of GFP+Gr-1+ cells in PB of primary and secondary recipients of empty vector or BCR-ABL–transduced BM cells from WT or HIF1α−/− donor mice (n = 5). Mean values (± SD) are shown. (D) The total numbers of GFP+Gr-1+ cells in PB of primary and secondary recipients of empty vector or BCR-ABL–transduced BM cells from WT or HIF1α−/− donor mice (n = 5). Mean values (± SD) are shown. *P < .05; **P < .01; ***P < .001. (E) The spleen weight of primary and secondary recipients of empty vector or BCR-ABL–transduced BM cells from WT or HIF1α−/− donor mice (n = 12). Mean values (± SD) are shown; ***P < .001. (F) Gross appearance of the lungs and spleens showed severe lung hemorrhages and splenomegaly in secondary recipients of WT LSCs but not HIF1α−/− LSCs at day 14 after BMT, compared with those from recipients of WT and HIF1α−/− HSCs. (G) H&E staining of tissue sections from lung and spleen of secondary recipients. The scale bar represents 50 μm. (H) Kaplan-Meier survival curves for the secondary BM transplantation. BM cells from WT or HIF1αflox/flox mice were transduced with BCR-ABL-iCre-GFP retrovirus, and transplanted into lethally irradiated recipient mice. CML cells without HIF1α failed to induce CML disease in the secondary BMT. (I) The percentages of leukemia cells in PB at day 14 in WT and HIF1α−/− secondary CML mice.

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