TT30 inhibits C3 fragment deposition on PNH RBCs in the EASA. (A-H) aNHS + TT30 3μM. (A-D) C3b/iC3b and C3d deposition at 1 hour. (E-H) C3b/iC3b and C3d deposition at 24 hours. (A-E) forward scatter (y-axis) versus side scatter (SSC). (B-F) CD59 APC (y-axis) versus C3b/iC3b FITC (x-axis; mAb 7C12). (C-G) CD59 APC (y-axis) versus C3d biotinylated-streptavidin PE (x-axis; mAb A702); (D-H) 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). (I-P) aNHS +3E7 2μM. (I-L) C3b/iC3b and C3d deposition at 1 hour. (M-P) C3b/iC3b and C3d deposition at 24 hours. (I-M) forward scatter (y-axis) versus SSC. (J-N) CD59 APC (y-axis) versus C3b/iC3b FITC (x-axis; mAb 7C12). (K-O) CD59 APC (y-axis) versus C3d biotinylated-streptavidin PE (x-axis; mAb A702); (L-P) 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 I-M).