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Figure 2. Non-reducing gel electrophoresis and chemotaxis assays of recombinant wild-type (WT) S100A8 and ala42S100A8 exposed to 10–5M hypochlorite (OCl). (A) Coomassie-blue-stained gel (1) WT S100A8 and (2) ala42S100A8. ala42S100A8 did not form covalently bound dimers. (B) Western blot analysis of WT S100A8 and mutated S100A8 with monoclonal antibodies to S100A8. (1) WT S100A8; (2) ala42S100A8. The antibodies recognized both WT S100A8 and ala42S100A8. (C) The data represent the average of 3 experiments performed in duplicate (± SD) with cells isolated from different adult volunteers. Transwell fugetaxis assays of neutrophils with WT S100A8 and ala42S100A8 oxidized by 10–5 M OCl on ice for 30 min. The fugetactic effect of ala42S100A8 was resistant to oxidation, whereas the WT S100A8 protein’s fugetactic effect was inhibited by oxidation (#p < 0.05; *p < 0.01 when compared with control with no compound added). We calculated the migration ratio by dividing the number of cells that migrated to the lower well in each experiment by the number of cells in the lower well in the untreated control wells.
