Figure 8. Expression and immunoprecipitation of wild-type or GDAY/AAAA mutant NPRA in recombinant HEK-293 cells.
Confluent recombinant HEK-293 cells expressing either wild-type or mutant receptors were labelled with [35S]methionine. Cell lysates (200 μg of protein) were immunoprecipitated using the site-directed polyclonal antibodies against NPRA. The labelled protein was separated on a 7.5% (w/v) polyacrylamide gel as stated in the Materials and methods section. Lanes a and b represent the expressed recombinant wild-type and GDAY/AAAA mutant receptor bands in HEK-293 cells respectively. The arrow indicates the position of the 135 kDa NPRA protein band. The positions of molecular-mass standards are shown on the right in kDa.