FIGURE 7.

PTH inhibits ectodomain cleavage by internalization of the PTHR. CHO cells stably expressing HA-PTHR were infected with adenovirus encoding for Dyn^WT or dynamin K44A (Dyn^K44A). 48 h after the infection, the cells were stimulated with 100 nm PTH(1–34) for 30 min or 12 h. A, cells were fixed, permeabilized, and stained with mouse anti-HA antibody followed by a Cy2-labeled anti-mouse antibody. HA-PTHR was visualized by confocal microscopy. B, cells were lysed, and the proteins were separated by SDS-PAGE and blotted to polyvinylidene difluoride membranes. PTHR was detected in Western blots with an anti-HA antibody. The positions of molecular weight standards are marked on the left. The arrowheads indicate cleaved and uncleaved PTHR species.