Creation of a genetic system for analysis of

Creation of a genetic system for analysis of the phagocyte respiratory burst: high-level reconstitution of the NADPH oxidase in a nonhematopoietic system by Marianne O. Price, Linda C. Mc. Phail, J. David Lambeth, Chang-Hoon Han, Ulla G. Knaus, and Mary C. Dinauer Blood Volume 99(8): 2653 -2661 April 15, 2002 © 2002 by American Society of Hematology

High-level superoxide production in transgenic COS-phox cells. Superoxide production by either wild-type COS-7 cells (▪) or transgenic COS-phox cells (■), following detachment from tissueculture dishes and stimulation with either 0. 4 μg/m. L PMA or 100 μM AA, . . . Marianne O. Price et al. Blood 2002; 99: 2653 -2661 © 2002 by American Society of Hematology

Effect of p 47 phox on serine residues 303, 304, and 379 NADPH oxidase activation in COS-phox cells. COS cells stably expressing gp 91 phox and p 22 phox were transiently transfected with wild -type p 67 phox plus either wild-type p 47 phox (p 47 wt) or p 47 phox mu. . . Marianne O. Price et al. Blood 2002; 99: 2653 -2661 © 2002 by American Society of Hematology

Effect of soluble agonists on membrane translocation of cytosolic NADPH oxidase components in COS-phoxcells. COS-phox cells were activated with 0. 4 μg PMA (10 minutes, 37°C) or 100 μM AA (5 minutes, 37°C), as indicated. Marianne O. Price et al. Blood 2002; 99: 2653 -2661 © 2002 by American Society of Hematology

Comparison of PKC isoform expression in COS-7 cells and human neutrophils. Membrane (Mb) and cytosolic (Cyt) fractions were isolated from whole-cell lysates of human neutrophils or transgenic COS-7 cells expressing gp 91 phox /p 22 phox. Marianne O. Price et al. Blood 2002; 99: 2653 -2661 © 2002 by American Society of Hematology

Effect of dominant-negative Rac or Rho. GDI expression on agonist-elicited superoxide production by COS-phoxcells. COS-phox cells were transiently transfected with expression vectors encoding dominant-negative Rac 1(T 17 N), Rac 2(T 17 N), or Rho. GDI. Marianne O. Price et al. Blood 2002; 99: 2653 -2661 © 2002 by American Society of Hematology

Effect of Val 204 Ala p 67 phox mutation on NADPH oxidase activation in whole cells. (A) COS-7 cells stably expressing gp 91 phox, p 22 phox, and p 47 phox, but not p 67 phox, were transiently transfected with expression vectors encoding either wild-type p 67 phox or p 67 p. . . Marianne O. Price et al. Blood 2002; 99: 2653 -2661 © 2002 by American Society of Hematology