Endogenous suppression of neutral-active and calcium-dependent phospholipase A2 in human polymorphonuclear leukocytes

Group II phospholipase A2 mRNA synthesis is stimulated by two distinct mechanisms in rat vascular smooth muscle cells

“…Phospholip~e activity was measured by the hy~olysis of ["Cloleic acid (Amersham)-Iabeled ~c~eric~ia coli phospholipids [24]. The assay mixture contained 100 mM Tris, pH 7.4, and 1 mM CaCl2.…”

“…The medium of the culture was diluted to produce hydrolysis of up to 10% of the substrate. Reaction mixtures were incubated for 1 h at 37"C, and the released ['4C]oleic acid was extracted and measured as described elsewhere [24].…”

Transforming growth factors type‐β and dexamethasone attenuate group II phospholipase A2 gene expression by interleukin‐1 and forskolin in rat mesangial cells

“…PLAz activity was detem~ined using [I -"'C]oleate-labelled E. co/i as substrate as described [28]. Assay mixtures (1.0 ml)contained 100 mM Tris-HCI (pH 7.4), I.0 mM CaCI:, 5 nM E. coli phospholipid (3000-SWOcpm) and the enzyme to be tested at a dilution producing approximately 5% substrate hydrolysis.…”

“…Assay mixtures (1.0 ml)contained 100 mM Tris-HCI (pH 7.4), I.0 mM CaCI:, 5 nM E. coli phospholipid (3000-SWOcpm) and the enzyme to be tested at a dilution producing approximately 5% substrate hydrolysis. Reactions were stopped after 1 h and the liberated ["C]olcate was extracted by a modified Dolt extraction procedure [28].…”

PDGF suppresses the activation of group II phospholipase A2gene expression by interleukin I and forskolin in mesangial cells

“…were \tdstlcd tu'lcc wrth PBS dud wzubalcd with I ml of RPM1 1640. contdmmp0 I !n@ml of fdttyacld-free bovmc serum dlbullrln 61ytm) PLA? actwlty was determined usmg [I-%]oleate-lnbelled C to/r d5 substrate as described [24] Assay mixtures (I 0 ml) contcuncd IO0 mM Tns-HCI (pH 7 4). 1 0 mM CaCI,, 5 nM C LO/I phosphohpld (3000-5000 cpm) and the enzyme to be tested nt d dllutlon producmg approxIm,~tely 5% substrate hydrolyslb Reactions were stopped after 1 h dnd the hberated ["Cloleate wds extracted by a modlhed Dole extrdctlon procedure [24] For mimunoprccrpltatlon.…”