Compound A inhibits IFN-γ promoter activity and cytokine production.

<p><i>A,</i> EL4 cells were cotransfected with 9 µg of IFN-γ promoter-driven luciferase plasmid (IFN-γ-Luc) plus 9 µg of GR expression vector and with 9 µg of T-bet expression vector. After 16 h in culture, cells were stimulated for 5 h with Compound A (CpdA, 10 µM). Results, as folds, normalized to β-galactosidase activity, are expressed as mean ± SEM (n = 6, * p<0.001 vs. basal without T-bet, ** p<0.001 vs. T-bet without CpdA), averaged from three independent experiments. <i>B,</i> Purification of T cells was achieved by a conventional technique involving cell adhesion to plastic and then to a nylon wool column and alternatevely by FACS sorting. Purified T cells were activated with PMA (P) and Ionomycin (I) during 24 h and then incubated with CpdA for 5 h. Supernatants were used to measure mouse IFN-γ according to the manufacturer’s instructions by ELISA. Results are expressed as mean ± SEM (n = 4, * p<0.001 vs. basal without CpdA, ** p<0.001 vs. P+I without CpdA) of one representative experiment of three independent experiments with similar results.