Fig. 5.

Analysis of gbp2 promoter chromatin and of gbp2 in Stat1^−/− cells. (A) STAT1-deficient fibroblasts were pretreated with dox for 6 h and then transiently transfected with pRETRO-tet-OFF-FLAG-IRF1. Twenty-four hours after transfection, dox was removed from the cells (−) or left on the cells (dox) for an additional 24 h. Control cells were treated with 10 ng/ml (240 units/ml) IFN-γ for 1 h. Formaldehyde-cross-linked chromatin was isolated and subjected to IP with IRF1 Abs. The proximal gbp2 promoter was analyzed by PCR. The specificity for the IP was determined by using preimmune serum (C) as negative control, and amplification of input DNA (IN) by PCR. (B) RNA was isolated, reverse-transcribed, and analyzed for endogenous gbp2 expression by real-time PCR.