Figure 8. Effect of sense and antisense PKC isoenzyme oligonucleotide transfections on NADPH oxidase subunit phosphorylation.

(A) After 48 h of transfection, cells were incubated with Hcy (100 μM) for 30 min and the levels of serine-phosphorylated p47^phox and p67^phox proteins were determined by immunoprecipitation and Western-blot analysis. Results were normalized to total p47^phox and p67^phox proteins in the immunoprecipitates. The relative densitometric units for the control levels were expressed as 100%. (B) Intracellular superoxide anion levels were measured by the NBT reduction assay after cells were transfected with sense or antisense PKC isoenzyme oligonucleotide for 48 h. The level of superoxide anions in control cells was expressed as 100%. Results are means±S.E.M. for three experiments each performed in duplicate. *Significantly different from the control value (P<0.05). ^#Significantly different from the value obtained from Hcy-treated cells (P<0.05).