He staining solution

T-2 toxin was purchased from Pribolab Pte. Ltd. (Singapore). BA was purchased from Sigma (St. Louis, MO, USA). VE, used as positive control [56 (link),57 (link)], was bought from Sigma-Aldrich (St Louis, MO, USA). MDA, GSH, SOD, GSH-PX and CAT assay kits were acquired from Nanjing Jiancheng Biotech (Nanjing, Jiangsu, China). The H&E staining solution was purchased from Servicebio (Wuhan, Hubei, China). IgG, SIgA, IgM, C3, C4 and DAO kits were supplied from Elabscience Biotechnology Co., Ltd. (Wuhan, Hubei, China). SYBR Green I fluorescent dyes and the Primescript RT reagent kit were from Takara (Shiga, Japan) and trizol was supplied by Life Technologies (Carlsbad, CA, USA). The IKB-α antibody, NF-κB p65 antibody and β-actin antibody were bought from Cell Signaling Technology, Inc. (Danvers, MA, USA), and all other reagents were analytical grade.

Chemicals are listed as follows: A804598 (HY-100483, MedChemExpress Co., Ltd. United States); IL-1Ra (HY-P7029A, MCE, United States); DMSO (D8370, Solarbio, China); Mouse NLRP3 ELISA kit (JYM0765Mo, Wuhan ColorfulGene biological technology Co., Ltd. China); Mouse IL-1β ELISA kit (JYM0531Mo, Wuhan ColorfulGene biological technology Co., Ltd. China); BCA Protein Assay Kit (PC0020, Solarbio, China); SDS-PAGE Gel Rapid Preparation Kit (G2037-50T, Servicebio, China); Recombinant Anti-beta Actin antibody (GB15003, Servicebio, China); NLRP3 Antibody (DF7438, Affinity Biologicals, Canada); Recombinant Anti-P2X7 antibody (ab259942, abcam, United Kingdom); HRP-conjugated goat anti-rabbit IgG (GB23204, Servicebio, China); Anti-Iba1 Rabbit pAb (GB11105, Servicebio, China); Cy3-labeled goat anti-rabbit IgG (GB21303, Servicebio, China); FITC-labeled goat anti-rabbit IgG (GB22303, Servicebio, China); Anti-BrdU Mouse mAb (GB12051, Servicebio, China); Anti-NeuN Rabbit pAb (GB11138, Servicebio, China); HE staining solution (G1005-100ML, Servicebio, China).

The lung, liver, and kidney samples of mice were fixed using fixative (Solarbio, China), embedded using HistoCore Arcadia (Leica, German) and cut into 4 μm-thick sections using RM2016 (Leica, German). HE and TUNEL staining was carried out using HE staining solution (Servicebio, China) and a TUNEL Detection Kit (Beyotime, China), respectively. The injury score was analyzed according to HE staining as described in a previous study (Li et al., 2022 (link)) by investigators who were unaware of treatment. Alveolar septum thickening, renal tubular epithelial cell swelling, inflammatory cell infiltration, hepatocyte swelling, focal necrosis, and diffuse coagulation were included in the manifestations of multiple organ injury. The injury areas were scored as follows: 0 for normal, 1 for <25% damage, 2 for 25–50% damage, 3 for 50–75% damage, 4 for 75–90% damage, and 5 for >90% damage.