Figure 4.

Red blood cells (RBCs) induce necroptosis of lung endothelial cells (ECs) and inhibition of necroptosis attenuates lung EC high-mobility group box 1 (HMGB1) release. (A) Viability of human lung microvascular ECs following RBC treatment, *P = 0.054. (B) HMGB1 release following RBC treatment, P = 0.03. necrostatin (Nec)-1–ZVAD significantly attenuates HMGB1 release with three of the six units tested, P ≤ 0.01. Data representative of three independent studies. (C) HMGB1 staining in lung EC 4 hours after RBC treatment or RBC treatment in the presence of Nec-1 (50 μM). red = HMGB1; blue = DAPI; green = F-actin. DAPI = 4′,6-diamidino-2-phenylindole.