Figure 9. Effect of LY333531 on PKC activity, NADPH oxidase subunit phosphorylation and superoxide anion levels in monocytes.
(A) Cells were incubated in the absence (Control) or presence of Hcy (100 μM) for 30 min. In some experiments, cells were pre-incubated with a PKC inhibitor (LY333531; 5 nM) for 30 min, followed by incubation with Hcy (100 μM) or without Hcy for another 30 min. PKC activity was measured. (B) Serine-phosphorylated p47phox and p67phox protein subunits were determined by immunoprecipitation and Western-blot analysis. Results are expressed as means±S.E.M. for three separate experiments. (C) Intracellular superoxide anion levels were measured by the NBT reduction assay. The level of superoxide anions in control cells was expressed as 100%. Results are expressed as means±S.E.M. for four separate experiments each performed in duplicate. The values obtained from control cells were expressed as an arbitrary unit of 100%. *Significantly different from the control value (P<0.05). #Significantly different from the value obtained from Hcy-treated cells (P<0.05).