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. Author manuscript; available in PMC: 2016 Nov 25.
Published in final edited form as: Thromb Haemost. 2015 Aug 13;114(6):1144–1155. doi: 10.1160/TH15-01-0079

Figure 3. Purification and APC binding to EPCR-GPI.

Figure 3

EPCR-GPI expressed in HEK-293 cells was purified from cell lysates. (A) Coomassie stain and (B) Western blot of purified EPCR-GPI under non-reducing conditions. (C) APC binding to recombinant soluble EPCR (sEPCR; circle) and EPCR-GPI (square). Signals were corrected for aspecific binding of APC to wells without EPCR. Data was fitted to a non-linear regression curve for one-site specific binding to calculate the Bmax and apparent KD, followed by normalized of the data to the corresponding Bmax. (D) Specificity of APC binding to EPCR-GPI using the blocking (rcr-252) and non-blocking (rcr-2) control anti-EPCR antibodies (both used at 20 μg/ml). (C-D) Shown are mean ± SD of at least three independent experiments. (E) Representative experiment of cell membrane fractionation on a density gradient of EPCR-GPI or wt-EPCR transfected HEK-293 cells. TfR denotes the transferrin receptor.