FIGURE 8.

Detection of a cleaved form of the PTHR in human kidney. Glycoproteins from human kidney tissue or from CHO cells stably expressing HA-PTHR, which had been treated for 12 h with 100 nm PTH(1–34), were enriched by wheat germ agglutinin affinity purification. Thereafter proteins were deglycosylated with PNGase F for 16 h. The enzyme reactions were stopped by the addition of SDS sample buffer, and the proteins were separated by SDS-PAGE and transferred to polyvinylidene difluoride membranes. PTHR was detected by Western blot using an anti-PTHR-CT antiserum. The arrowheads indicate cleaved and uncleaved PTHR species.