Figure 3. Quantification of the affinity of GST-MIM-S1 for MIM.
The lysate of cells expressing MIM-Myc (A) or MIM-I-BAR-Myc (B) were incubated with GST-MIM-S1 at the concentrations as indicated. The GST-MIM-S1 binding proteins were isolated by pull-down. The amounts of unbound MIM-Myc or MIM-I-BAR-Myc proteins in the supernatants after precipitation were measured by Western blot using anti-Myc.As the loading control, β-actin in the supernatants was also measured. The data was analyzed by Prism software, which predicted Kd values of 110 nM and 65 nM for MIM-Myc and MIM-I-BAR-Myc, respectively.