Fig. 2.

Functional leucine-rich repeat G protein-coupled receptors (LGRs) in hepatocytes. A: LGR4 immunoreactivity in hepatocytes. Isolated hepatocytes were stained with LGR4 antibody (green), and nuclei were stained with DAPI (blue). B: translocation of LGR4 upon treatment with R-spondin-1 (Rspo1). Isolated hepatocytes were incubated with Rspo1 (200 ng/ml) or saline for 60 min, and membrane and cytoplasm proteins were isolated and used for Western blot to detect LGR4 translocation; β-actin was used as loading control. C: internalization of LGR4. Isolated hepatocytes were incubated with Rspo1 (200 ng/ml) for the time indicated and then stained with LGR4 antibody (green) to detect mobilization of LGR4 receptor upon activation by Rspo1. Each experiment was repeated 3 times.