Fig. 4
From: The DISC1 R264Q variant increases affinity for the dopamine D2 receptor and increases GSK3 activity
The mouse DISC1-L100P mutation increases the binding affinity between DISC1 and D2R. a. GST-DISC1NT and GST-Disc1-L100PNT affinity “pull-down” of D2R from mouse striatal tissue. Various concentrations (50, 100, 150 μg) of GST-DISC1NT or GST-Disc1-L100PNT were incubated with mouse striatal extract and the precipitated proteins were immunoblotted with anti-D2R. b. Densitometric analysis of precipitated D2R. The intensity of D2R was quantified by densitometry (software: ImageLab, Bio-Rad). GST-DISC1NT compared with GST-Disc1-L100PNT by two-way ANOVA followed by Bonferroni post hoc test (**p < 0.01 as compared to that of 100 μg GST-DISC1NT, ###p < 0.001 as compared to that of 150 μg GST-DISC1NT, n = 3). Data was shown as mean ± SEM. c-d. Surface Plasmon Resonance (SPR) analysis of the concentration-dependent binding of GST-DISC1NT (c) or GST-Disc1-L100PNT(d) with immobilized TAT-D2pep[K211-T225] peptide. Binding affinity was measured using a range of concentrations from 25 nM to 400 nM for GST-Disc1-L100PNT, and 100 nM to 1.6 μM for GST-DISC1NT, respectively. The duration of the association and dissociation phases was set to 180 s, 300 s, respectively. Flow rate was set to 50 uL/min to reduce the effect of mass transport limitations. Regeneration of the sensor chip was achieved by adding 10 mM NaOH, 1 M NaCl