Fig 7. Recombinant PN expressed in HEK293VKOR cells has the expected pIs of non- γ-carboxylated protein.
Purified PN0, 2.3 μg produced in the presence of 10 μg/mL vitamin K by HEK293VKOR cells was run on two-dimensional isoelectric focusing/electrophoresis and Western blotting gels. The parallel 8% gels were transferred to PVDF membranes and probed with (A) mouse anti-PN, or (B) mouse anti-Gla. Isoelectric points of PN positive spots ranged between pH 7.0 and > pH 8.0, consistent with unmodified protein. No anti-Gla-positive spots were detected.