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Figure 1


Fig. 1. Analysis of NFAT mRNA expression in human neutrophils. Total RNA was extracted from highly purified unstimulated neutrophils from both human healthy donors (N1-N3) and human allergic patients (N4-N6), as well as from PBLs (L) and Jurkat T cells (J) stimulated with 100 nM PMA plus 500 nM ionomycin, and from highly purified eosinophils (E) used as positive and negative controls. (A,B) RNA samples were analysed by conventional RT-PCR, using specific primers for the amplification of the indicated NFAT isoform transcripts (A), or of the Charcot-Leyden crystal protein (C-L) transcripts (B). Results from amplification of the beta2-microglobulin (beta2m) housekeeping gene transcript are shown in both panels. (C) RNA was isolated from neutrophils from allergic patients (AP) or healthy donors (HD) or from PBLs (L). The levels of NFAT1, NFAT2, NFAT4 and NFAT5 mRNAs were analysed by real-time PCR analysis as indicated in the Materials and Methods section. The levels of mRNAs were standardised against the level of beta2-microglobulin (beta2m) mRNA. Values are mean ± s.e.m. from eight individuals in each group.





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