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. 2020 Apr;105(4):e180–e185. doi: 10.3324/haematol.2019.232587

Figure 2.

Figure 2.

Transient expression of wild-type and mutant coagulation factor V proteins in COS-1 cells. Radiolabelled coagulation factor V (FV) was immunoprecipitated from cell lysates (A) and conditioned media (B) of COS-1 cells transfected with plasmids expressing FV (wild-type or mutant). Mock-transfected COS-1 cells represent the negative control (lane M). Proteins were electrophoresed on 4% SDS-PAGE gels under non-reducing conditions. Arrowheads indicate FV monomeric molecules. Antigen (Ag) levels of recombinant FV were measured both in cell extracts (C) and in the corresponding conditioned media (D) by an ELISA assay. FV activity (Fx) was measured in conditioned media by a functional assay based on the prothrombin time (E). In all graphs, bars represent means + standard deviations of at least two independent experiments, each performed in duplicate. The mean value of wild-type FV was set as 100%. Significance levels of t-tests are shown: *P<0.05, **P<0.01, ***P<0.001. Details on all methodological aspects can be found in9.