FIGURE 5.

KLF6 interacts with p65 in the nucleus. A, KLF6 interacts with p65 in the nucleus. HEK293 cells (5 × 10⁷) were left untreated or treated with TNFα (20 ng/ml) or IL-1β (10 ng/ml) for the indicated time points. Cells fractionation was performed, and the cytoplasmic and nuclear proteins were prepared in 1 ml of homogenization buffer or nuclear extraction buffer, respectively, following by coimmunoprecipitation and immunoblot analysis. B, IκBα but not p50 disrupts the p65-KLF6 interaction in a dose-dependent manner. HEK293 cells (3 × 10⁶) were transfected with the indicated plasmids together with IκBα (1 or 2 μg) or p50 (1 or 2 μg) expression plasmids. Coimmunoprecipitation and immunoblots were performed with the indicated antibodies. C, KLF6 does not affect p65-IκBα interaction. HEK293 cells (3 × 10⁶) were transfected with the indicated plasmids together with KLF6-expression plasmids (4 or 8 μg). Coimmunoprecipitation and immunoblots were performed with the indicated antibodies.