Figure 3.

p97 binds selectively to RanBP2 in the presence of RanGTP. A blot overlay approach was used to analyze the binding of p97 to proteins of salt-washed NE (swNE, 2 OD₂₆₀, lanes 1–3), or to purified RanBP2 (10 ng, lanes 4–5). Protein samples were separated by SDS-PAGE on a 5–15% gradient gel, electrophoretically transferred to nitrocellulose, and the nitrocellulose membrane was renatured. Nitrocellulose strips were incubated with GSTp97, RanGDP, and RanGTP as indicated. After cross-linking, bound p97 was detected using anti-GST antibodies.