Figure 5

Protein expression analysis. A. ELISA for selected Cytokines and Chemokines. Cytokine and chemokine generation/release was determined from control-unstimulated mature mast cell (Basal); following IgE-sensitization (IgE) for 24 hours; and FcεRI aggregation by addition of the anti-human IgE to sensitized cells for 24 hours (XLFcεRI). Cell culture supernatants were analyzed for bioactive IL-1β; IL-6; IL-8; MCP-3 and RANTES by ELISA. Results shown are the mean plus the standard deviation of triplicate measurements and are representative of four separate experiments. B. Western Blot analysis of SPHK1. Upper panel; differential expression of SPHK1 was assessed by Western blot on lysates from control unstimulated cells (Basal); following IgE-sensitization (IgE) for 24 hours; and FcεRI aggregation by addition of the anti-human IgE to sensitized cells for 24 hours (XLFcεRI). Lower panel; the blots were probed for α-tubulin (control for equal loading). Results shown are representative of four separate experiments.