Fig. 5

Coagulation occurred after parasite egress from iRBCs. a–c (i) Western blotting analysis of coagulation. a “Med” and “Sup” indicate PDS- or NCS-containing medium and PDS- or NCS-cultured supernatant, respectively. b After the addition of protease inhibitors (PMSF phenylmethylsulfonyl fluoride, DE dabigatran etexilate). c “Lysate” indicates the isolated parasite lysate prepared using the precipitation method. The isolated parasite lysate prepared from iRBCs cultured in NCS-containing medium was added to plasma. (i) The absence of fibrinogen indicated coagulation, save for in the isolated parasite lysate (lanes 3 and 4). (ii) Visual confirmation of coagulation. Tubes, shown in lanes 3, 4, 5, and 6 in (i), containing 100 µL of the reaction mixture were rotated at a 90° angle onto their sides after completion of the coagulation reaction. All experiments were performed in triplicate and representative data are shown