Figure 7.

Individual real time PCR analysis confirmed the inhibition of fluticasone-induced gene expression by activated mast cell conditioned media. Healthy ASM cells were pre-treated with control mast cell media, conditioned media from 24-h non-activated, or 24-h activated mast cells for 24 h before cells were washed and stimulated with or without 100 nM fluticasone for an additional 6 h. Expression of anti-inflammatory genes of GILZ (A), MKP-1 (B), FKBP5 (C), and PIK3R1 (D) was assessed by real-time PCR. Data were normalized to the housekeeping gene (GAPDH) and presented as fold change over the basal condition using the 2^−ΔΔct method as Means ± SEM, n = 7 different cell lines (*p < 0.05, **p < 0.01). (E) Validation of the gene array data using individual PCR. The log2 fold induction of the 5 genes shows a strong correlation with the results found using Qiagen RT2 profiling gene array and qRT-PCR, (R² = 0.99, P = 0.0003).