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Manipulating the onset of cell cycle withdrawal in differentiated erythroid cells with cyclin-dependent kinases and inhibitors by Igor Matushansky, Farshid.

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Presentation on theme: "Manipulating the onset of cell cycle withdrawal in differentiated erythroid cells with cyclin-dependent kinases and inhibitors by Igor Matushansky, Farshid."— Presentation transcript:

1 Manipulating the onset of cell cycle withdrawal in differentiated erythroid cells with cyclin-dependent kinases and inhibitors by Igor Matushansky, Farshid Radparvar, and Arthur I. Skoultchi Blood Volume 96(8):2755-2764 October 15, 2000 ©2000 by American Society of Hematology

2 Changes in CDK activities during erythroleukemia cell terminal differentiation.Logarithmically growing MEL cells were induced to differentiate with 5 mmol/L HMBA as described in “Materials and methods.” (A) At the indicated times the cell number (○) and the... Igor Matushansky et al. Blood 2000;96:2755-2764 ©2000 by American Society of Hematology

3 Levels of cyclins and CDK inhibitors and their associations with CDKs during MEL cell differentiation.Total cellular protein extracts were prepared at the indicated times of HMBA treatment, and the levels of (A) specific cyclins and (C) CDK inhibitors were... Igor Matushansky et al. Blood 2000;96:2755-2764 ©2000 by American Society of Hematology

4 Characterization of p18 and p27 MEL cell transfectants.(A) MEL cell transfectant clones expressing either p18 or p27 under control of the tetracycline inducible promoter in pUHD 10-3 were cultured either in the absence (−) or presence (+) of 1 μg/mL Dox for... Igor Matushansky et al. Blood 2000;96:2755-2764 ©2000 by American Society of Hematology

5 Effect of p18 and p27 on proliferation of differentiated cells.We cultured p18 and p27 MEL cell transfectants in the presence of 5 mmol/L HMBA for 12 hours. Igor Matushansky et al. Blood 2000;96:2755-2764 ©2000 by American Society of Hematology

6 Inducible expression of exogenous CDK2, CDK4R24C, and CDK6R31C in MEL cell transfectants.(A, B) The extracts were prepared from the indicated CDK-HA MEL cell transfectants cultured in the absence (−) or presence (+) of Dox for 36 hours. Igor Matushansky et al. Blood 2000;96:2755-2764 ©2000 by American Society of Hematology

7 Comparison of exogenous and endogenous kinase activities in undifferentiated and differentiated MEL cell CDK transfectants.The indicated MEL cell transfectants were cultured in the absence (0) or presence (120) of HMBA for 120 hours and then further culture... Igor Matushansky et al. Blood 2000;96:2755-2764 ©2000 by American Society of Hematology

8 Synergistic activities of CDK2 and CDK4 in causing extensive proliferation of fully differentiated cells.(A) The indicated MEL cell CDK transfectants were cultured in the presence of 5 mmol/L HMBA for 48 hours and then plated in plasma clots either in the a... Igor Matushansky et al. Blood 2000;96:2755-2764 ©2000 by American Society of Hematology


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