Fig 2: Nec-1 pretreatment inhibits intestinal cell necroptosis in LPS-challenged piglets.A The representative jejunal ultrastructural images by TEM. LPS-induced necrotic ultrastructural changes such as nuclear deformation, nuclear membrane rupture and chromatin overflowing D, and Nec-1 reduced the necrotic ultrastructural changes. Original magnification: 5000 ×, scale bars = 1 μm. B Localization of RIP1 and HMGB1 by immunohistochemistry staining. Original magnification: 400 ×, scale bars = 22.4 μm. C–K Protein expression of necroptosis-related signals by western blotting, and serum HMGB1 content by ELISA. Cy HMGB1, cytoplasmic HMGB1. Nu HMGB1, nuclear HMGB1. The pigs were pretreated intraperitoneally with Nec-1 at 1.0 mg/kg body weight or equal volume of 2% DMSO solution 30 min before the intraperitoneal injection of LPS or saline, and the pigs were sacrificed at 4 h after LPS or saline injection. Values are means ± SE, n = 7. abcMeans without a common letter differ, p < 0.05.

Fig 3: LPS challenge dynamically induces muscle necroptosis in weanling pigs. Longissimus dorsi muscle samples were collected from the control group (0 h after injection of saline) and LPS-treated groups (1, 2, 4, 8,12, and 24 h after injection of LPS). (A–C) mRNA levels of RIP1, RIP3, and MLKL. (D–M) Protein expression of t-RIP1, p-RIP1, t-RIP3, p-RIP3, t-MLKL, and p-MLKL. Data are presented as means ± SEM, n = 6. *** p < 0.001, ** p < 0.01, * p < 0.05, and # 0.05 < p < 0.10, different from the control group (0 h).