Comparison of the alternative pathway–mediated hemolysis between the C3-sufficient and C3-Dpl human or C3-deficient rat sera. For the measurements of alternative pathway–mediated hemolysis, E^rabb cells were incubated with different concentrations of NHS ranging from 5% to 40%, C3-Dpl human sera (A) or WT C3-deficient (C3KO) rat sera (B) in GVB-Mg⁺⁺/EGTA buffer, and hemolysis was quantitated by measuring levels of released hemoglobin in the supernatants. (C) In some experiments, E^rabb cells were incubated with 100% human or rat sera, and hemolysis was assessed by following the same protocol. These data show almost complete abolition of hemolysis in the C3-Dpl human sera and C3-deficient rat sera. *P < .05.