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. 2020 Jul 14;295(35):12498–12511. doi: 10.1074/jbc.RA120.013692

Figure 1.

Figure 1.

PT is the most abundant RAGE-interactor detected in human plasma by a VC1 pulldown assay. A, SDS-PAGE of the fractions from pulldown assays carried out in 20 mm HEPES, 100 mm NaCl, 0.5% Triton X-100, pH 7.1. IN, input; UB, unbound fraction; B, bound material obtained after boiling the beads for 5 min in SDS sample buffer. The arrowhead indicates the 75-kDa polypeptide identified as PT and present in the VC1-STV beads and absent in the control (STV beads). VC1 and STV are also indicated. Proteins were stained by Coomassie Blue. B, Western blotting (WB) analysis using anti-PT mAb of fractions from a VC1 pulldown assay. C, Western blotting analysis of a VC1 pulldown assay using anti-Gla domain mAb. B2/3 and B1/3 indicate that 2/3 and 1/3 of the total bound material were loaded. D, VC1 pulldown assay using purified PT instead of plasma. The indicated samples were analyzed by SDS-PAGE and proteins were stained by Coomassie Blue.