Figure 1.

Stat3-C protein is successfully overexpressed in mouse liver and activates Stat3 DNA binding. (a) Anti-Stat3 and anti-FLAG immunoblots were performed to determine total and exogenous Stat3 protein in liver tissue, respectively, 3 days after adenovirus infection. (b) Immunohistochemical study using anti-Stat3 showed that Stat3-C protein is expressed in nuclei and cytoplasm of the liver in Stat3-C mice. Original magnification, ×200. (c) Electrophoretic mobility shift assay (EMSA) analysis showed elevated Stat3 DNA binding in the liver of Stat3-C mice. Five micrograms of nuclear extract was incubated with SIE-m67 probe. In order to confirm specific Stat3 DNA binding, competition (comp.) and supershift assays were performed using cold SIE-m67 probe and anti-Stat3. (d) Western blot analysis of Stat3 protein in various tissues of control and LS3-KO mice. These data are representative of at least three independent experiments. FP, free probe; ss, supershift; IB, immunoblot.