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. 2000 Jul 4;97(15):8525–8529. doi: 10.1073/pnas.150149097

Figure 1.

Figure 1

Cleavage of pro-ANP by corin. (a) Western analysis of recombinant corin. Human embryonic kidney 293 cells were stably transfected with corin expression vector pcDNACorin/V5 or a control vector pcDNA. Conditioned medium, total cell lysate, and membrane fractions were prepared based on methods described previously (27). Expression of corin was detected by SDS/PAGE and Western blotting using an anti-V5 antibody. (b) Cotransfection of pro-ANP and corin expression vectors. Pro-ANP expression vector pcDNAproANP was cotransfected into 293 cells with vectors expressing corin (pcDNACorin, lane 2), prothrombin (pProthrombin, lane 3), hepsin (pHepsin, lane 4), or a control vector (pcDNA, lane 1). Conditioned medium was collected and processing of pro-ANP was analyzed by Western blotting using an anti-V5 antibody. (c) Incubation of conditioned medium containing pro-ANP with transfected cells. Conditioned medium from pcDNAproANP transfected cells was collected and incubated with 293 cells transfected with vectors expressing corin (pcDNACorin, lane 2), prothrombin (pProthrombin, lane 3), hepsin (pHepsin, lane 4), or a control vector (pcDNA, lane 1). Processing of pro-ANP was analyzed by Western blotting.