Figure 4

Induction of CEACAM1 expression in MCF7 cells with IFN γ. A. Top, RT-PCR with total RNA isolated from MCF7 cells, either untreated (-) or treated for 6 h with 500 U/ml of IFN γ (+). Bottom, CEACAM1 mRNA levels were monitored by real time PCR and normalized to GAPDH (triplicates ± SD). B. Total cellular protein lysates from MDA-MB-468, MCF10A and MCF7 cells either untreated (-) or treated (+) with 500 u/ml IFN γ were subjected to Western blot and probed with antibodies to IRF1, IRF2, and CEACAM1. β-actin was used as a loading control. Closed and open circle indicate two CEACAM1 isoforms with a different migration from MDA-MB-468 and MCF10A cells, respectively. C. Chromatin immunoprecipitation of CEACAM1 promoter DNA from MCF7 cells untreated (-) or treated with 500 u/ml of IFN γ for 6 h. Antibodies to USF1, USF2, IRF1 and a control IgG were used, numbers refer to percent of input.