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BCR-ABL and Interleukin 3 Promote Haematopoietic Cell Proliferation and Survival through Modulation of Cyclin D2 and p27Kip1 Expression

Parada, Yelena, Banerji, Lolita, Glassford, Janet, Lea, Nicholas C., Collado, Manuel, Rivas, Carmen, Lewis, John L., Gordon, Myrtle Y., Thomas, N. Shaun B., Lam, Eric W.-F.·2001·Journal of Biological Chemistry
DOI 10.1074/jbc.m101885200
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FIG. 1 Effects of IL-3 and/or BCRABL on cell cycle progression and survival of BaF3 cells. A, cell cycle analysis of BaF3 and BaF3-p210 cells in the absence or presence of IL-3 ( upper panel ). BaF3 and BaF3-p210 cells were incubated for 24 h in the absence or presence of IL-3. The resultant cells were collected, permeablized, and stained with propidium iodide. The cell cycle profile was expressed as 'number of cells' against 'DNA content' and the percentages of cells in each phases ( , G1, G1 , S, and G2/M) were indicated. Cells undergoing apoptosis was represented by those with , G1 DNA content. B, cell cycle analysis of BaF3 cells after IL-3 withdrawal ( lower panel ). Cycling BaF3 cells previously cultured in IL-3-supplemented medium were incubated in IL-3-free growth media and sampled at 0, 6, 12, 24, 48, and 72 h for cell cycle analysis as described above.
FIG. 1 Effects of IL-3 and/or BCRABL on cell cycle progression and survival of BaF3 cells. A, cell cycle analysis of BaF3 and BaF3-p210 cells in the absence or presence of IL-3 ( upper panel ). BaF3 and BaF3-p210 cells were incubated for 24 h in the absence or presence of IL-3. The resultant cells were collected, permeablized, and stained with propidium iodide. The cell cycle profile was expressed as 'number of cells' against 'DNA content' and the percentages of cells in each phases ( , G1, G1 , S, and G2/M) were indicated. Cells undergoing apoptosis was represented by those with , G1 DNA content. B, cell cycle analysis of BaF3 cells after IL-3 withdrawal ( lower panel ). Cycling BaF3 cells previously cultured in IL-3-supplemented medium were incubated in IL-3-free growth media and sampled at 0, 6, 12, 24, 48, and 72 h for cell cycle analysis as described above.CC-BY · From the PDF
Image 2
CC-BY · From the PDF
FIG. 2 Expression of cell cycle regulators in BaF3 cells in the absence and presence of IL-3 and/or BCR-ABL. Cell lysates extracted from BaF3 and BaF3-p210 cultured with and without IL-3 for 24 h, as described in the legend to Fig. 1 A , were Western blotted for BCR-ABL, cyclin D2, -D3, -E, and -A, CDK2, -4, and -6, CDC2, pRB, p107, p130, p16 INK4a , and p21 Cip1 expression using the respective antibodies and for CDK4/6 and CDK2 activity using the phospho-pRB antibodies. Total RNAs from BaF3 and BaF3-p210 cells 1 IL-3 were also extracted for Northern blot analysis for expression of cyclin D2, cyclin D3, and p27 Kip1 mRNA ( lower right panel ). Equal loading of total RNAs was confirmed by staining with ethidium bromide.
FIG. 2 Expression of cell cycle regulators in BaF3 cells in the absence and presence of IL-3 and/or BCR-ABL. Cell lysates extracted from BaF3 and BaF3-p210 cultured with and without IL-3 for 24 h, as described in the legend to Fig. 1 A , were Western blotted for BCR-ABL, cyclin D2, -D3, -E, and -A, CDK2, -4, and -6, CDC2, pRB, p107, p130, p16 INK4a , and p21 Cip1 expression using the respective antibodies and for CDK4/6 and CDK2 activity using the phospho-pRB antibodies. Total RNAs from BaF3 and BaF3-p210 cells 1 IL-3 were also extracted for Northern blot analysis for expression of cyclin D2, cyclin D3, and p27 Kip1 mRNA ( lower right panel ). Equal loading of total RNAs was confirmed by staining with ethidium bromide.CC-BY · From the PDF
FIG. 3 Expression of cell cycle regulators in BaF3 cells after IL-3 deprivation. Lysates derived from BaF3 cells at 0, 6, 12, 24, 48 and 72 h after IL-3 withdrawal were immunoblotted for cyclin D2, -D3, and -E, CDK2, -4, and -6, pRB, p107, p130, and p27 Kip1 .
FIG. 3 Expression of cell cycle regulators in BaF3 cells after IL-3 deprivation. Lysates derived from BaF3 cells at 0, 6, 12, 24, 48 and 72 h after IL-3 withdrawal were immunoblotted for cyclin D2, -D3, and -E, CDK2, -4, and -6, pRB, p107, p130, and p27 Kip1 .CC-BY · From the PDF
Image 5
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Image 6
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Image 7
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FIG. 6 Effects of STI571 on the expression of cell cycle regulators in BaF3 cells expressing BCR-ABL. A, BaF3-p210 cells were incubated with or without STI571 6 IL-3 for 24 h. The expression of cyclin D2, -D3, and -E, CDK2, -4, and -6, pRB, p107, p130, p27 Kip1 , and p21 Cip1 , and CDK4/6 and CDK2 activity were detected by immunoblotting. B , the expression of cyclin D2, cyclin D3, and p27 Kip1 mRNA was also studied by Northern blotting ( right panel ) in these cells.
FIG. 6 Effects of STI571 on the expression of cell cycle regulators in BaF3 cells expressing BCR-ABL. A, BaF3-p210 cells were incubated with or without STI571 6 IL-3 for 24 h. The expression of cyclin D2, -D3, and -E, CDK2, -4, and -6, pRB, p107, p130, p27 Kip1 , and p21 Cip1 , and CDK4/6 and CDK2 activity were detected by immunoblotting. B , the expression of cyclin D2, cyclin D3, and p27 Kip1 mRNA was also studied by Northern blotting ( right panel ) in these cells.CC-BY · From the PDF
Image 9
CC-BY · From the PDF
Image 10
CC-BY · From the PDF
FIG. 8 Effects of overexpression of cyclin D2 on cell cycle progression and survival of BaF3 and BaF3-p210 cells. A, cell cycle profile of BaF3 and BaF3-cyclin D2 cells after IL-3 deprivation for 0, 24, 48, and 72 h. B, cell cycle profile of BaF3-p210 and BaF3-p210 cells overexpressing cyclin D2 after STI571 treatment for 0, 24, and 48 h. C, the expression of cyclin D2 in the BaF3-p210 and BaF3-p210-cyclin D2 cells after STI571 treatment for 0, 24, and 48 h was analyzed by Western blotting.
FIG. 8 Effects of overexpression of cyclin D2 on cell cycle progression and survival of BaF3 and BaF3-p210 cells. A, cell cycle profile of BaF3 and BaF3-cyclin D2 cells after IL-3 deprivation for 0, 24, 48, and 72 h. B, cell cycle profile of BaF3-p210 and BaF3-p210 cells overexpressing cyclin D2 after STI571 treatment for 0, 24, and 48 h. C, the expression of cyclin D2 in the BaF3-p210 and BaF3-p210-cyclin D2 cells after STI571 treatment for 0, 24, and 48 h was analyzed by Western blotting.CC-BY · From the PDF
Image 12
CC-BY · From the PDF
Image 13
CC-BY · From the PDF
FIG. 9 Effects of overexpression of cyclin D2 on the expression of downstream cell cycle regulators in BaF3 cells. Cell lysates from control BaF3 and BaF3-cyclin D2 serum-starved cells after different times were Western blotted for the expression of cyclin D2, -D3, and -E, CDK2, -4, and -6, p27 Kip1 , pRB, p107, p130, and E2F1, and activity of CDK4/6 and CDK2.
FIG. 9 Effects of overexpression of cyclin D2 on the expression of downstream cell cycle regulators in BaF3 cells. Cell lysates from control BaF3 and BaF3-cyclin D2 serum-starved cells after different times were Western blotted for the expression of cyclin D2, -D3, and -E, CDK2, -4, and -6, p27 Kip1 , pRB, p107, p130, and E2F1, and activity of CDK4/6 and CDK2.CC-BY · From the PDF

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Metadata

Authors
Parada, Y., Banerji, L., Glassford, J., Lea, N. C., Collado, M., Rivas, C., Lewis, J. L., Gordon, M. Y., Thomas, N. S. B., & Lam, E. W.-F.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2001-06
Volume
276
Issue
26
Pages
23572-23580
ISSN
0021-9258
DOI
10.1074/jbc.m101885200
Cited by
88
References
0
Language
en
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CoChat. (2026). Citation check: BCR-ABL and Interleukin 3 Promote Haematopoietic Cell Proliferation and Survival through Modulation of Cyclin D2 and p27Kip1 Expression. https://app.cochat.ai/papers/10.1074/jbc.m101885200
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Parada, Y., Banerji, L., Glassford, J., Lea, N. C., Collado, M., Rivas, C., Lewis, J. L., Gordon, M. Y., Thomas, N. S. B., & Lam, E. W.-F. (2001). BCR-ABL and Interleukin 3 Promote Haematopoietic Cell Proliferation and Survival through Modulation of Cyclin D2 and p27Kip1 Expression. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.m101885200