Figure 6. Comparison of the GST pull-down of TFII-I in NE by wild-type and mutant Gal1 polypeptides.
The GST proteins were incubated overnight with glutathione beads at 4 °C in 60% buffer D. After washing to remove the unbound material, the beads were then incubated with NE for 12 hours at 4 °C. Material bound to the various beads was eluted with glutathione (16 mM) and analyzed by SDS-PAGE and immunoblotting. Panel A: immunoblotting with anti-TFII-I (# 558). Panel B:immunoblotting with anti-GST to ascertain that approximately equal amounts of GST proteins were bound to the beads in the pull-down assay.