FIG 8.

PI3K and Akt activities are required for PIP₃-induced phagocytosis of nonmotile P. aeruginosa. (A) Murine BMDCs were coincubated with PA14 motAB motCD bacteria for 45 min after pretreatment of cells in the absence or presence of 12 μM PIP₃. Following intracellular staining for phospho-Akt, Akt activation was quantified by FACS analyses. Mean fluorescence intensity (MFI) was normalized to untreated cells (set as 1, y axis). (B and C) Phagocytic uptake of PA14 motAB motCD (MOI of 10) by gentamicin protection assay with murine WT BMDCs pretreated with 12 μM PIP₃ with or without 100 μM LY-294002, 5 μM Akt inhibitor VIII, or 10 μM cytochalasin D, as indicated. The data were normalized to phagocytosis of PA14 motAB motCD without PIP₃ treatment and are represented as fold increase in phagocytosis. Data were analyzed using one-way ANOVA with Tukey's post hoc analyses and are representative of at least two independent biological experiments (A, n = 4; B and C, n = 8). ***, P ≤ 0.0005; ns, not significant.