Figure 1. .

Fresh and aged plasmas trigger oxidative burst and protein phosphorylation in neutrophils. (A) Representative results of ROS generation in untreated normal human neutrophils (Control), and neutrophils treated for 1 h with fresh (1 day preparation) non-leukocyte-reduced plasma (NLR-1D), aged (42 day preparation) non-leukocyte-reduced plasma (NLR-42D), aged leukocyte-reduced plasma (LR-42D), and NLR-42D plus NADPH oxidase inhibitor DPI (NLR-42D + DPI). The results are expressed as means ± SD from 3 experiments. fMLP: formyl-Met-Leu-Phe (as a positive control). (*p < 0.05, compared with NLR-42D). (B) Western blot analysis of protein tyrosine phosphorylation in response to different preparations of plasma. Normal human neutrophils were incubated with the different plasma preparations for 1 h. Whole cell lysates were blotted with anti-pY20 antibody. (C) Normal human neutrophils were incubated with the different plasma preparations for 1 hr. Whole cell lysates prepared from these neutrophils were subjected to antibody array analysis with immunoblotted with anti-pY20-HRP. (D-E) Western blot analysis demonstrated tyrosine phosphorylation of IKK, p105 and p50 in response to plasma treatment (*p < 0.05, compared with NLR-42D). 1D represents the ratio of p-IKK value over IKK. 1E represents the ratio of p105 value over p50.