Figure 2
Kinetics of C3 fragment deposition and processing in the EASA. (A-D) Fresh PNH RBCs from untreated patient, baseline data. (A) Forward scatter (y-axis) versus side scatter (SSC). (B) CD59 APC (y-axis) versus C3b/iC3b FITC (x-axis; mAb 7C12). (C) CD59 APC (y-axis) versus C3d biotinylated-streptavidin PE (x-axis; mAb A702); (D) C3d biotinylated-streptavidin PE (y-axis) versus C3b/iC3b FITC (x-axis; mAb 3E7). Red and blue dots represent intact RBCs and RBC ghosts, respectively, as gated based on physical parameters (panels A-E). (E-L) aNHS. (E-H) C3b/iC3b and C3d deposition at 1 hour. (I-L) C3b/iC3b and C3d deposition at 24 hours. (E-I) forward scatter (y-axis) versus SSC. (F-J) CD59 APC (y-axis) versus C3b/iC3b FITC (x-axis; mAb 7C12). (G-K) CD59 APC (y-axis) versus C3d biotinylated-streptavidin PE (x-axis; mAb A702); (H-L) C3d biotinylated-streptavidin PE (y-axis) versus C3b/iC3b FITC (x-axis; mAb 3E7). Red and blue dots represent intact RBCs and RBC ghosts, respectively, as gated based on physical parameters (panels E-I). (M-T) aEcuHS. (M-P) C3b/iC3b and C3d deposition at 1 hour. (Q-T) C3b/iC3b and C3d deposition at 24 hours. (M-Q) forward scatter (y-axis) versus SSC. (N-R) CD59 APC (y-axis) versus C3b/iC3b FITC (x-axis; mAb 7C12). (O-S) CD59 APC (y-axis) versus C3d biotinylated-streptavidin PE (x-axis; mAb A702); P, T: C3d biotinylated-Streptavidin PE (y-axis) versus C3b/iC3b FITC (x-axis; mAb 3E7). Red and blue dots represent intact RBCs and RBC ghosts, respectively, as gated based on physical parameters (panels M-P).

Kinetics of C3 fragment deposition and processing in the EASA. (A-D) Fresh PNH RBCs from untreated patient, baseline data. (A) Forward scatter (y-axis) versus side scatter (SSC). (B) CD59 APC (y-axis) versus C3b/iC3b FITC (x-axis; mAb 7C12). (C) CD59 APC (y-axis) versus C3d biotinylated-streptavidin PE (x-axis; mAb A702); (D) C3d biotinylated-streptavidin PE (y-axis) versus C3b/iC3b FITC (x-axis; mAb 3E7). Red and blue dots represent intact RBCs and RBC ghosts, respectively, as gated based on physical parameters (panels A-E). (E-L) aNHS. (E-H) C3b/iC3b and C3d deposition at 1 hour. (I-L) C3b/iC3b and C3d deposition at 24 hours. (E-I) forward scatter (y-axis) versus SSC. (F-J) CD59 APC (y-axis) versus C3b/iC3b FITC (x-axis; mAb 7C12). (G-K) CD59 APC (y-axis) versus C3d biotinylated-streptavidin PE (x-axis; mAb A702); (H-L) C3d biotinylated-streptavidin PE (y-axis) versus C3b/iC3b FITC (x-axis; mAb 3E7). Red and blue dots represent intact RBCs and RBC ghosts, respectively, as gated based on physical parameters (panels E-I). (M-T) aEcuHS. (M-P) C3b/iC3b and C3d deposition at 1 hour. (Q-T) C3b/iC3b and C3d deposition at 24 hours. (M-Q) forward scatter (y-axis) versus SSC. (N-R) CD59 APC (y-axis) versus C3b/iC3b FITC (x-axis; mAb 7C12). (O-S) CD59 APC (y-axis) versus C3d biotinylated-streptavidin PE (x-axis; mAb A702); P, T: C3d biotinylated-Streptavidin PE (y-axis) versus C3b/iC3b FITC (x-axis; mAb 3E7). Red and blue dots represent intact RBCs and RBC ghosts, respectively, as gated based on physical parameters (panels M-P).

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