Survey
* Your assessment is very important for improving the work of artificial intelligence, which forms the content of this project
Download Powerpoint - Blood Journal
Document related concepts
no text concepts found
Transcript
Transduction of a dominant-negative H-Ras into human eosinophils attenuates extracellular signal–regulated kinase activation and interleukin-5–mediated cell viability by David J. Hall, Jin Cui, Mary Ellen Bates, Barbara A. Stout, Leo Koenderman, Paul J. Coffer, and Paul J. Bertics Blood Volume 98(7):2014-2021 October 1, 2001 ©2001 by American Society of Hematology Tat-dominant-negative N 17 Ras construct.The dominant-negative N17 H-Ras gene was cloned into the pTat-HA vector as described in “Materials and methods.” Six His residues precede the 11-amino acid Tat peptide. David J. Hall et al. Blood 2001;98:2014-2021 ©2001 by American Society of Hematology FITC-labeled Tat-dn-H-Ras enters eosinophils.Tat-dn-H-Ras was labeled with FITC, as described in “Materials and methods.” Human blood eosinophils resuspended in pH 6.8 HBSS without calcium or magnesium and 1% fetal calf serum were incubated with 100 nM FITC... David J. Hall et al. Blood 2001;98:2014-2021 ©2001 by American Society of Hematology Transduced Tat-dn-H-Ras blocks activation of endogenous Ras on IL-5 stimulation.Human blood eosinophils were incubated with 100 nM Tat-dn-H-Ras for 30 minutes and then stimulated for 7 minutes with 1 nM IL-5. David J. Hall et al. Blood 2001;98:2014-2021 ©2001 by American Society of Hematology Dosage of Tat-dn-H-Ras affects IL-5–stimulated ERK 1 and ERK 2 activation.(A) Human blood eosinophils were incubated with various concentrations of Tat-dn-H-Ras for 30 minutes and then stimulated for 7 minutes with 1 nM IL-5. David J. Hall et al. Blood 2001;98:2014-2021 ©2001 by American Society of Hematology Time course of Tat-dn-H-Ras incubation as a function of inhibition of ERK 1 and ERK 2 activation.(A) Human eosinophils were incubated with 100 nM Tat-dn-H-Ras for different time intervals and then stimulated for 7 minutes with 1 nM IL-5. David J. Hall et al. Blood 2001;98:2014-2021 ©2001 by American Society of Hematology Phosphorylation of the ERK substrate p90 Rsk1 is suppressed by Tat-dn-H-Ras.Human blood eosinophils were incubated with 100 nM Tat-dn-H-Ras and then stimulated for 7 minutes with 1 nM IL-5. David J. Hall et al. Blood 2001;98:2014-2021 ©2001 by American Society of Hematology STAT5 phosphorylation after IL-5 stimulation is not blocked by Tat-dn-H-Ras.Human blood eosinophils were incubated with 100 nM of Tat-dn-H-Ras or Tat-GFP for 30 minutes and then stimulated for 15 minutes with 300 pM IL-5. David J. Hall et al. Blood 2001;98:2014-2021 ©2001 by American Society of Hematology Effect of Tat-dn-H-Ras and MEK inhibitor U0126 on eosinophil viability.Blood eosinophils were incubated with no IL-5 (▧), 10 pM IL-5 (▪), 100 nM Tat-dn-H-Ras (■), 100 nM Tat-GFP (░), 10 μM U0126 (▩), or 50 μM PD98059 (▤) for up to 60 hours. David J. Hall et al. Blood 2001;98:2014-2021 ©2001 by American Society of Hematology
Related documents