Fig. 12. GST pull-down of GKLF by Sp1.
The GST pull-down experiments are described under “Experimental Procedures.” Western blot analysis was performed on various protein fractions using a polyclonal rabbit GKLF antibody (13). Lanes 1 and 2 contained input lysates from mock- (vector alone-) transformed and GKLF-transformed bacteria, respectively. Lanes 3–5 contained the flow-through (F.T.) fractions of GKLF that did not bind to the glutathione-Sepharose 4B beads containing GST, GST-Sp1ZnF, and GST-Sp1Q1, respectively. Lanes 6–8 contained the eluant (E.T.) fractions of GKLF that bound to the beads and were eluted with 1 mm reduced glutathione. The location of GKLF is indicated and has a molecular mass of 24 kDa. The asterisk denotes a nonspecific cross-reacting material that was present in the lysates of mock-transformed bacteria.