For the in vitro GST pull - down assay, 20 μL of glutathione -
Sepharose 4B beads (Amersham Pharmacia Biotech) with immobilized GST or GST / Sp1 fusion proteins (4 μg each) were incubated with HuH - 7 whole - cell extracts (500 μg) at 4 °C for 6 hours.