In vivo reconstitution of AP activity in MASP-1/3−/− mice with rMASP-3

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In vivo reconstitution of AP activity in MASP-1/3−/− mice with rMASP-3. In vivo reconstitution of AP activity in MASP-1/3−/− mice with rMASP-3. An ex vivo assessment of the AP by lysis of rabbit blood erythrocytes (RBEs) and cleavage of proDF by Western blot analysis at 24 h from MASP-1/3−/−–injected mice with active (rM3) or inactive or mutated rM3 (rM3 S/A). (A) Natural RBEs lysis by WT serum using various dilutions as expected. (B) No RBEs lysis by MASP-1/3−/− mouse serum reconstituted with rM3 S/A. (C) RBEs lysis by MASP-1/3−/− serum reconstituted with rM3. (D) Western blot showing the cleavage of proFD into FD only after injecting rM3, not rM3 S/A (lane 7 versus lane 5). Data from MASP-1/3−/− mice shown (B and C) as mean OD ± SEM. All samples were run in duplicate with highly reproducible data from two MASP-1/3−/− male and two female mice. Western blot were repeated three times. M1/3 + rM3, MASP-1/3−/− mice treated active rMASP-3; M1/3 + rM3 S/A, MASP-1/3−/− mice treated inactive rMASP-3; dil, serum dilutions; Tx, treatment. Nirmal K. Banda et al. ImmunoHorizons 2018;2:274-295 Copyright © 2018 The Authors