Figure 2

Phagocyte functions are impaired in PI3Kγ-/- mice. Peritoneal macrophages of three wildtype and three knockout littermates were isolated at different time points, cells were pooled and used for in vitro experiments. (A) Production of NO, IL-1β, and IL-6 was measured after 24 hours culture without restimulation. Mean and SEM of triplicates are shown for one of two experiments. < det: below detection limit (10 pg/ml) (B) Macrophages isolated at day 2 of AIA were starved for 4 hours, restimulated for 5 min with C5a, as indicated, lysed and subjected to immunoblotting for Akt, phospho-Akt (pAkt), and β-actin. (C) Macrophages isolated at day 2 of AIA were seeded into transwell plates and migrated towards different concentrations of C5a for 20 hours. Afterwards, membranes were washed and cells at the lower side were stained and counted microscopically. One of two experiments is shown (mean and SEM). (D) Joint sections at day 2 of AIA were prepared and stained with Mac-1 (macrophages, neutrophils) or Gr-1 (neutrophils) specific antibodies (upper panel) and positive cells were counted (lower panel, mean and SEM of 6 animals). *: statistically significant difference, p < 0.05.