Stable expression of Recombinant EP
in HEK293 cells. (A) Western blot analysis for over-expressed EP1 receptor in HEK293 cells. HEK293 cells transfected with cDNA of EP1 receptor (lane 1) or untransfected (lane 2) were solubilized and separated by 10% SDS-PAGE, and then transferred to a nitrocellulose membrane. The expressed protein was stained with an antibody against EP1. (B) Immunofluorescence micrographs of HEK293 cells. The general procedures for the indirect immunostaining are described in the methods section. In brief, the EP1 receptor stable cell line (Lane 1) or untransfected HEK293 controls (Lane 2) were grown on cover-slides. The cells were generally permeabilized by 1% saponin and then incubated with the anti-EP1 polyclonal rabbit antibody. The bound antibodies were stained by FITC-labeled goat anti-rabbit IgG. The stained cells were examined by fluorescence microscopy.