Smooth muscle cell migration and proliferation are mediated by distinct phases of activation of the intracellular messenger mitogen-activated protein.

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Presentation transcript:

Smooth muscle cell migration and proliferation are mediated by distinct phases of activation of the intracellular messenger mitogen-activated protein kinase Peter R. Nelson, MD, Shinji Yamamura, MD, Leila Mureebe, MD, Hiroyuki Itoh, MD, PhD, K.Craig Kent, MD, FACS Journal of Vascular Surgery Volume 27, Issue 1, Pages 117-125 (January 1998) DOI: 10.1016/S0741-5214(98)70298-8 Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Effect of PD098059 on PDGF-BB–induced activation of MAPK. Quiescent SMCs were simultaneously incubated with varying concentrations of PD098059 and PDGF-BB (5 ng/ml) for 10 minutes. MAPK activity, as measured by phosphorylation of its substrate myelin basic protein, was determined using immunoprecipitation and SDS-PAGE techniques as described in Methods. This experiment was repeated three times using cells derived from different saphenous veins. A representative example is displayed. Journal of Vascular Surgery 1998 27, 117-125DOI: (10.1016/S0741-5214(98)70298-8) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Effect of PD098059 SMC chemotaxis. Quiescent SMCs were stimulated with PDGF (5 ng/ml) in the presence of increasing concentrations of PD098059. Chemotaxis was determined using a 4-hour microchemotaxis chamber assay as described in Methods. Results are expressed as mean percent of control (PDGF-stimulated) chemotaxis ± SD. All experiments were repeated three times using cells derived from different saphenous veins. A representative example is displayed. Journal of Vascular Surgery 1998 27, 117-125DOI: (10.1016/S0741-5214(98)70298-8) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Effect of PD098059 on SMC proliferation. Quiescent SMCs were stimulated with PDGF (5 ng/ml) in the presence of increasing concentrations of PD098059. SMC proliferation was determined by measuring (A) DNA synthesis by 3H-thymidine incorporation, or (B) cell counts as described in Methods. Results are expressed as mean percent of control (PDGF-stimulated) proliferation ± SD. All experiments were repeated three times using cells derived from different saphenous veins. A representative example is displayed. Journal of Vascular Surgery 1998 27, 117-125DOI: (10.1016/S0741-5214(98)70298-8) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Relationship between temporal activation of MAPK and SMC migration and proliferation. A, DNA synthesis was measured (as described in Methods) with PD098059 (50 μmol/L) added either simultaneously or at intervals after PDGF stimulation (5 ng/ml). Results are expressed as mean percent of control (PDGF-stimulated) proliferation ± SD. B, Microchemotaxis assays were performed (as described in Methods) with addition of PD098059 (50 μmol/L) simultaneously or at intervals after exposure of SMCs to PDGF (5 ng/ml). Results are expressed as mean percent of control (PDGF-stimulated) chemotaxis ± SD. All experiments were repeated three times using cells derived from different saphenous veins. Representative examples are displayed. Journal of Vascular Surgery 1998 27, 117-125DOI: (10.1016/S0741-5214(98)70298-8) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions