Fig. 2.
Displacement binding assay of WT and ICL1 PROKR2 mutants. To measure PROK2 binding by displacement analysis, COS7 cells transfected with either WT, R80C, R85C, or R85H PROKR2 were incubated with 125I-MIT-1 in the presence of increasing concentrations (10−10 to 10−6 m) of unlabeled PROK2 for 15 min, after which cells were washed and radioactivity assayed in cell lysates. The PROK2 displacement curves were used to calculate Bmax and Kd for each PROKR2 receptor. Results are expressed as percentage of the WT PROKR2 maximal binding. Each point is the mean ± se of three independent experiments.