Figure 3.
A, GST pulldown assay. 50 pmol of GST-RHINO(45–64) (lanes 7–9) or GST alone (lanes 4–6) was immobilized on GSH-Sepharose 4B beads and incubated with 100 pmol of purified WT 9ΔC-1-1 (WT) or its derivatives, m1 (K155A/R244A/Q254A in RAD1) and m2 (F64A/M256A/F266A in RAD1). Input 9-1-1 (20 fmol, lanes 1–3) and bound proteins (lanes 1–9) were analyzed by immunoblotting. B, the intensity of the HUS1 band in each lane was quantified, and the amount of bound HUS1 was plotted. The pulldown assay was performed twice, and results of quantification were shown (exp.#1 and exp.#2).