FIGURE 4.

Detection of PTH-stimulated adenylyl cyclase and ERK1/2 phosphorylation. HEK293 cells were transiently transfected with WT-P1R, Flag-P1R-L, and Flag-P1R-P. After 48 h transfection, the cells were incubated with vehicle (0.1% BSA) or PTH (100 nM). (Upper) cAMP accumulation was measured as described in Materials and Methods. Data were summarized as the mean ± se of three independent experiments. (Lower) A representative immunoblot showed that PTH stimulated ERK1/2 phosphorylation (Lower, upper panel). The amount of ERK1/2 expression in each sample was used as an internal loading control (Lower, lower panel).