Fig. 4. Storage in AS-5 leads to increased RBC necroptosis. Fresh human RBCs or those stored in AS-5 for 1 week were treated with A.) VLY, B.) ILY, C.) PLY, and D.) LLO. Storage in AS-5 for 1 week leads to increased RBC necroptosis by all toxins relative to fresh RBCs. RBC necroptosis after 1 week of storage is inhibited by nec-1s, GSK'872, and NSA. Hemolysis values were normalized to basal RBC death in the absence of toxin treatment. Results shown are from 3 independent experiments. Two-way ANOVA, ***p<0.001.