Fig. 5.

Expression and ligand-independent activity human GPR30 activity with and without PSD-95 in yeast S. cerevisiae. (A) CY2797 and EY-1 yeast strains transformed with empty plasmid (Mock), plasmid containing GPR30, or plasmids containing GPR30 and PSD-95 were immunoblotted with GPR30 and pan–membrane-associated guanylate kinase (MAGUK) Ab. (B, C) Cell growth was followed for various times and graphed as OD₆₀₀. Two-way ANOVA with Tukey’s multiple comparisons test was used to statistically analyze the relationship between all groups. (D) Cells were treated with vehicle (−) or 1 μM G-1 (+), and OD₆₀₀ was determined and graphed for time 0 hours and 30 hours of treatment. Statistical analysis was performed using two-way ANOVA with Tukey’s multiple comparisons test. (E) Data from (C) were normalized to the OD₆₀₀ at t=0 and analyzed using repeated measures one-way ANOVA with test for trend to assess time-dependent linear relationships for each condition. The results are either representative (A) or the means ± S.D. (C)–(E) of at least three independent experiments. In (A), molecular mass standards are indicated (left side). *P < 0.05; ****P < 0.0001; n.s., not significant.