Activation of PKB is not required for the migration of MoDCs in response to CCL19 and CCL21. MoDCs matured in the presence of PGE2 were incubated for 30 minutes with graded doses of (A) wortmannin or (B) Ly-294002 and were tested for migration to CCL21 and CCL19 in a transwell assay. Error bars indicate SEM. (C) To confirm the effectiveness of wortmannin, MoDCs treated with or without wortmannin were stimulated with CCL19 or CCL21 for 2 minutes. Whole-cell lysates were separated on SDS-PAGE, and activated PKB or Erk-1/2 was detected with an antibody against PKB phosphorylated at S473 (p-PKB) or Erk-1/2 dually phosphorylated at Thr202 and Tyr204 (p-Erk). Membranes were reprobed with an antibody reacting with total Erk-1/2 (t-Erk) to confirm equal protein loading. Results from 1 of 3 representative experiments using different MoDC preparations are shown.