Fig. 8. Involvement of NFAT in IgE-dependent COX2 expression in human neutrophils. Neutrophils from an allergic patient sensitised to G₃ antigen were pretreated with CsA or cell-permeable VIVIT, at the indicated doses for 1 hour, and then incubated for 24 hours with 1 µg/ml LPS or 10 µg/ml anti-IgE Ab (-IgE; A) or 10 µg/ml G₃ (B). COX2 mRNA levels were analysed by conventional RT-PCR, using COX2- and GAPDH-specific primers; COX2 and -actin protein expression was analysed by western blotting, and the levels of PGE₂ were measured in the culture medium supernatants, using an enzyme immunoassay. The values shown are the mean ± s.e.m. from three independent assays in which each measurement was performed in triplicate. All the experiments were performed at least in triplicate.