Time kinetic of B7-H1 expression and B7-H1 promoter targeting upon combined cytokine treatment with IL-4 and TNF α. A) Hal31RC was treated with either IL-4 or TNFα alone and with the combination of the two cytokines. B7-H1 protein expression was analyzed at the time points indicated by flow cytometry. Shown are fold increase in B7-H1 expression compared to untreated cells. Representative data of at least 3 different experiments are shown. B) B7-H1 mRNA expression in Hal31RC upon treatment with either IL-4 or TNFα alone and with the combination of the two cytokines for the time points indicated are shown. Using a B7-H1 plasmid as standard, B7-H1 transcript levels per 10.000 transcripts β-actin are shown. Data are obtained by real time PCR. Representative data of 3 different experiments are shown. C) BUF1088Mel was transfected with the mCherry B7-H1 promoter construct and stimulated with IL-4 and TNFα. The mCherry fluorescence was measured after 72 hrs by FACS and a representative histogram plot is depicted. The bar graph shows the combined result of a total of 3 independent assays.