Phorbol ester up-regulates capacities for nuclear
translocation and phosphorylation of 5-lipoxygenase in
Mono Mac 6 cells and human polymorphonuclear
leukocytes
by Oliver Werz, Jenny Klemm, Bengt Samuelsson, and Olof Rådmark
Blood
Volume 97(8):2487-2495
April 15, 2001
©2001 by American Society of Hematology
Translocation of 5-LO and up-regulation of 5-LO product formation in MM6 cells.(A) Western blot
analysis of 5-LO in subcellular fractions from MM6 cells.
Oliver Werz et al. Blood 2001;97:2487-2495
©2001 by American Society of Hematology
Dose response of PMA and mezerein for 5-LO product formation and 5-LO translocation in MM6
cells.Intact MM6 cells (3 × 106 in 1 mL PGC buffer) were primed for 10 minutes at 37°C with the
indicated concentrations of PMA (●) or mezerein (○).
Oliver Werz et al. Blood 2001;97:2487-2495
©2001 by American Society of Hematology
Dose response of A23187 regarding 5-LO product formation and 5-LO translocation in
PMNLs.For determination of 5-LO product formation in intact cells (A), human PMNLs (5 × 106 in
1 mL PGC buffer) were preincubated in the absence (●) or presence of 100 nM PMA...
Oliver Werz et al. Blood 2001;97:2487-2495
©2001 by American Society of Hematology
Dose response of PMA and mezerein for priming of 5-LO product formation and 5-LO
translocation in PMNLs.(A) To determine the effect on 5-LO product formation, PMNLs (5 × 106
cells in 1 mL PGC buffer) were primed with the indicated concentrations of PMA (●) ...
Oliver Werz et al. Blood 2001;97:2487-2495
©2001 by American Society of Hematology
Calphostin C inhibits 5-LO product formation and 5-LO redistribution in MM6 cells and
PMNLs.To determine the effects on 5-LO product formation or translocation, the indicated cell
numbers (in 1 mL PGC buffer) were incubated with the indicated concentrations...
Oliver Werz et al. Blood 2001;97:2487-2495
©2001 by American Society of Hematology
PMA up-regulates 5-LO phosphorylation by MK2 in MM6 cells: inhibition by calphostin
C.Phosphorylation of 5-LO by immunoprecipitates of MK2 from MM6 cells was determined by in
vitro kinase assays.
Oliver Werz et al. Blood 2001;97:2487-2495
©2001 by American Society of Hematology
PMA up-regulates 5-LO kinase activation in PMNLs: inhibition by calphostin C.PMNLs (5 × 107 in
1 mL PGC buffer) were preincubated at 37°C in the presence or absence of calphostin C, as
indicated, for 30 minutes under ordinary fluorescent light.
Oliver Werz et al. Blood 2001;97:2487-2495
©2001 by American Society of Hematology
[3H]arachidonic acid release in MM6 cells and human PMNLs.MM6 cells or freshly isolated
human PMNLs (2 × 106/mL in RPMI 1640 medium) were prelabeled with 0.25 μCi/mL [3H]AA for
90 minutes at 37°C and 5% CO2.
Oliver Werz et al. Blood 2001;97:2487-2495
©2001 by American Society of Hematology