Pin1 is required for TNF-α–induced priming of ROS production. (A) Neutrophils were incubated in Hanks buffer containing juglone (250nM) for 30 minutes; then TNF-α (20 ng/mL) was added for 20 minutes before stimulation with fMLF (10−7M). ROS production was measured with a luminol-amplified chemiluminescence technique. (B) Total chemiluminescence in each experimental condition is expressed as mean plus or minus SEM of 6 experiments. (C) PPIn (50 μM) was tested in the same conditions as juglone. (D) Data are mean plus or minus SEM of 6 experiments. (E) Neutrophils were incubated with juglone or PPIn and then stimulated with PMA (100 ng/mL) before measuring ROS production with a luminol-amplified chemiluminescence technique (one experiment representative of 3). (F) Total chemiluminescence in each experimental condition is expressed as mean plus or minus SEM of 3 experiments. *P < .01 compared with inhibitor-free conditions.
