Fig. 7.

KLF family proteins have the ability to homo- and heterodimerize via their DNA-binding domain. a–c GST pulldown assays were carried out using bacterially expressed GST, or the indicated GST-tagged proteins, and in vitro translated either full-length HA-TIEG1 (a, b) or GFP-TIEG1(DBD) (c). Autoradiographs are presented in the upper panels, whereas Coomassie brilliant blue stained SDS-polyacrylamide gels are shown in the lower panels. c, d TIEG1 homodimerization is not mediated by nucleic acids in the reaction mixture. GFP-TIEG1(DBD) was in vitro translated and divided into two. One half was used directly for GST pulldown assays (c, right panel), and the remaining half was treated with 250 units of benzonase for 20 min at room temperature prior to GST pulldown assays (c, left panel). d The benzonase used in the assay was active. The expression construct, GFP-TIEG1(DBD), was either treated with benzonase or left untreated, and the reaction analyzed on a 0.7% agarose gel followed by ethdium bromide staining. Data shown are representative of two independent experiments