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. 2007 Mar 8;104(12):5044–5049. doi: 10.1073/pnas.0611608104

Fig. 7.

Fig. 7.

The effect of IL-1 on the induction and activation of IRF-1 by IFN-γ. (A) EMSA of GAS–STAT complexes. WISH cells were treated for the indicated times with the indicated combinations of IFN-γ (100 units/ml), IL-1Ra (10 μg/ml), and IL-1α (2 ng/ml). Nuclear extracts were prepared and reacted with a GAS probe. (B) Immunoblots of IRF-1. WISH cells were treated with 2 ng/ml IL-1β for 1 h, followed by 100 units/ml IFN-γ. Total cell extract was obtained at the indicated times after IFN-γ addition and subjected to SDS/PAGE and immunoblotting with IRF-1 and β-actin antibodies. (C) EMSA of IRF-1–ISRE complexes. WISH cells were treated with 2 ng/ml IL-1β for 1 h, followed by 100 units/ml IFN-γ for 30 min. Total cell extract was subjected to EMSA with an ISRE probe. The identity of the complex was confirmed by supershifting with the indicated antibodies. The arrowhead identifies the ISRE complex, and the arrow identifies the supershifted complexes.