Activity of intracellular signaling pathways does not predict migratory capacity or cytokine secretion by DCs. MoDCs were activated using intact E coli and inhibitors added (as in Figure 2) 20 to 40 minutes prior to E coli exposure, and cells were harvested at 2 and 24 hours later, washed, and resuspended in lysis buffer. Strength of phosphorylation of (A) ERK1/2, (B) p38K, (C) PTEN, (D) unphosphorylated (active) PTEN, and (E) Akt-1 was analyzed. Activation of these pathways was assessed by quantitative immunoblotting. Data shown as means ± SD (ERK-1/2 and p38K at 2 hours from n = 9 and 24 hours from n = 5-6; all other proteins, n = 4 separate donors). *P < .05; **P < .01 compared with activation without inhibitors. (F) PC-PLC activity was measured in cellular lysates harvested 24 hours after activation (means ± SD, n = 6-8 separate donors), **P < .01 compared with activation without inhibitors (E coli). (G) Western blot analysis of MoDCs 2 hours following activation. Phosphorylation of Akt-1 is representative of n = 4 separate donors. p38K analysis acts as protein loading control.