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
Identification of CD9 extracellular domains important in regulation of CHO cell adhesion to fibronectin and fibronectin pericellular matrix assembly by George A. Cook, Celia M. Longhurst, Svetozar Grgurevich, Shila Cholera, Joseph T. Crossno, and Lisa K. Jennings Blood Volume 100(13):4502-4511 December 15, 2002 ©2002 by American Society of Hematology Flow cytometry analysis of CD9 cDNA-transfected CHO cells using anti-CD9 EC1 and EC2 antibodies.Cell suspensions were incubated with rabbit IgG (rIgG), anti-CD9 EC2 antibodies mAb7 or RAP5a, or anti-CD9 EC1 RAP2. George A. Cook et al. Blood 2002;100:4502-4511 ©2002 by American Society of Hematology CD9 influence on CHO cell adhesion to FN and pericellular FN matrix assembly is reversed by CD9 EC2 deletion mutant Δ173-192, Δ152-192, and Δ133-192 expression.(A) MOCK, A6, and CD9 deletion mutants Δ173-192, Δ152-192, and Δ133-192 CHO cells were allowed to... George A. Cook et al. Blood 2002;100:4502-4511 ©2002 by American Society of Hematology CD9 EC2 peptides 5b (135K-V172) and 6a (168P-I185) competitively block anti-CD9 mAb7 binding to soluble and cell surface CD9 as well as reverse the CD9 inhibitory influence on CHO cell adhesion to fibronectin.(A) Platelet lysate was added to either anti-CD9... George A. Cook et al. Blood 2002;100:4502-4511 ©2002 by American Society of Hematology Laser scanning confocal microscopy analysis revealed CD9 colocalized with integrin α5β1, but not all integrin subunit β1, on the basal surface of subconfluent FN-adherent CHO A6 cells.Cell cycle–synchronized CHO A6 cells were grown on FN-coated slides for 3... George A. Cook et al. Blood 2002;100:4502-4511 ©2002 by American Society of Hematology CD9 coimmunoprecipitates with integrin subunit β1 from CHO A6 but not CHO Δ133-192 cell lysates.Biotinylated surface proteins from CHO cell lysates were immunoprecipitated with antiCD9 mAb7 or RAP2, anti-β1 7E2, nonspecific binding mouse IgG, or anti-GPIb ... George A. Cook et al. Blood 2002;100:4502-4511 ©2002 by American Society of Hematology CD9 and cytoskeletal F-actin are colocalized on the basal surface of subconfluent FN-adherent CHO A6 cells.Cell cycle synchronized CHO A6 cells were grown on FN-coated slides for 3 hours, as described in “Materials and methods.” Laser scanning confocal micr... George A. Cook et al. Blood 2002;100:4502-4511 ©2002 by American Society of Hematology Truncation of CD9 EC2 reverses CD9-induced reduction of integrin α5β1 and cytoskeletal F-actin colocalization in fibronectin-adherent CHO cells.CHO MOCK, A6, and Δ133-192 cells were grown on FN for 3 hours, followed by integrin α5β1 and cytoskeleton F-actin... George A. Cook et al. Blood 2002;100:4502-4511 ©2002 by American Society of Hematology Truncation of CD9 EC2 reverses CD9 influence on spatial distribution of FAK as well as reduction of FAK and cytoskeletal F-actin colocalization in FN-adherent CHO cells.CHO MOCK, A6, and Δ133-192 cells were grown on FN for 3 hours, followed by FAK and cytos... George A. Cook et al. Blood 2002;100:4502-4511 ©2002 by American Society of Hematology Truncation of CD9 EC2 reverses CD9 influence on level of α-actinin expression and cytoskeletal F-actin colocalization in FN-adherent CHO cells.CHO MOCK, A6, and Δ133-228 cells were grown on FN for 3 hours, followed by α-actinin and cytoskeleton F-actin labe... George A. Cook et al. Blood 2002;100:4502-4511 ©2002 by American Society of Hematology Schematic model of CD9 and proposed functional domains.Amino acid sequence determination and antibody-binding studies suggest that CD9 consists of 2 EC loops, EC1 and EC2, and 4 TM domains (TM1-TM4), with the N and C termini located intracellularly.1 EC2 aa... George A. Cook et al. Blood 2002;100:4502-4511 ©2002 by American Society of Hematology
Related documents