Previous studies demonstrated that the intraperitoneal injection of epidermal growth factor (EGF) into mice resulted in the appearance, within minutes, of several tyrosine-phosphorylated proteins in liver nuclei. Two of these proteins have been identified as the transcription factors p91/p84 (Stat1 alpha/1 beta) (Ruff-Jamison, S., Chen, K., and Cohen, S. (1993) Science 261, 1733-1736). We have now identified, by Western blotting and immunoprecipitation, an additional EGF-modulated transcription factor, Stat3. We find that Stat3 is tyrosine-phosphorylated and present in mouse liver nuclei following either EGF or lipopolysaccharide administration. Gel shift analyses show that Stat3 is capable of specifically binding the SIE (a DNA sequence present in the c-fos promoter). Three active SIE binding complexes (SIF A, B, and C) exist in the nucleus after the administration of EGF: one complex that contains Stat3, one that contains Stat1, and a third complex that appears to contain both proteins. Only one active SIE binding complex, containing Stat3, was detected after the administration of lipopolysaccharide.