Wortmannin-sensitive activation of PKCζ is an early event in LPS-induced signal transduction.

(A, B) LPS stimulates the phosphorylation of PKCζ on the PDK1 target residue T410; quiescent BAC-1.2F5 cells were stimulated with 1.5 μg/mL LPS for the indicated time points. The presence of phosphorylated PKCζ was assessed by immunoblot analysis of either whole cell extracts (A) or anti-PKCζ immunoprecipitates (B). (C) Wortmannin affects LPS-stimulated PKCζ phosphorylation. BAC-1.2F5 cells were incubated with wortmannin (WM, 100 nmol/L, 20 minutes) before a 15-minute stimulation with 1.5 μg/mL LPS. Anti-PKCζ blots of whole cell extracts (A, C) or immunoprecipitates (B) are shown as a loading control.