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. 2012 Feb 15;16(4):329–338. doi: 10.1089/ars.2011.4043

FIG. 2.

FIG. 2.

Expression and subcellular location of VKORC1 and its homologues in HEK293 cells. (A) Western blot detection of the expression of VKORC1 and its bacterial homologues in HEK293 cells. HPC4-tagged proteins were transiently expressed in HEK293 cells for 48 h. Cell lysates were directly used for SDS-NuPAGE and protein bands were transferred to PVDF membrane. HPC4-tagged protein bands were probed with anti-HPC4 monoclonal antibody and visualized by ECL Western blot reagents. Lane 1, VKORC1-Y139F; lane 2, Mt-VKORH; lane 3, Rs-VKORH; lane 4, Ss-VKORH; M, marker, kDa. (B) Subcellular localization of VKORC1 and its bacterial homologues in HEK293 cells. GFP-tagged proteins were transiently expressed in HEK293 cells for 48 h. Cell images were collected from emission at 493–530 nm after excitation at 488 nm. GFP, green fluorescence protein; HPC4, peptide epitope from human protein C comprising residues EDQVDPRLIDGK; Rs-VKORH, Roseiflexus sp. RS-1 vitamin K epoxide reductase homologue; SDS-NuPAGE, Sodium dodecyl sulfate neutral polyacrylamide gel electrophoresis; Ss-VKORH, Synechococcus sp. vitamin K epoxide reductase homologue.