Recombinant murine il 1β 211 11b

Lipopolysaccharide (LPS, L4391-1MG), L-glutamic acid (G8415), and a hydrogen peroxide solution (30%w/w, H1009) were sourced from Sigma-Aldrich (United States). Recombinant murine IL-1β (211-11B) was procured from PEPROTECH (United States). Fluoxetine HCl (S27345, purity >98%), was obtained from Shanghai Yuanye Bio-Technology Co., Ltd. Hydroxytyrosol (purity >98%) was synthesized and purified by the Biopharmaceutical Laboratory of Shandong Academy of Pharmaceutical Sciences (China). Detailed structural and chromatographic information for HT is shown in Supplementary Figure S1.

Nec-1 was purchased from Selleck Chemicals (Houston, TX, United States). Nec-1 was dissolved in DMSO for in vitro use, and was dissolved in 5% DMSO, 45% PEG300 and ddH₂O, in turn. Recombinant murine IL-1β (#211-11B) was obtained from PeproTech (Rocky Hill, NJ, United States). Cell Counting Kit-8 (CCK-8), mouse antibody anti-GAPDH (BM3876), and secondary antibodies were purchased from Boster (Wuhan, China). Antibodies against RIPK1 (#3493), p-ERK (#4370), p-p38 (#4511), p-JNK (#9255), p-IkBa (#2859), p-p65 (#3033) and cleaved caspase-3 (#9964) were purchased from Cell Signaling Technology (Beverly, MA, United States). Rabbit antibodies against ERK (16443-1-AP), p38 (14064-1-AP), JNK (24164-1-AP), IkBa (10268-1-AP), p65 (10745-1-AP), matrix metalloproteinase (MMP)3 (17873-1-AP), TLR4 (19811-1-AP), HMGB1 (10829-1-AP), and stromal cell-derived factor-1 (SDF-1) (17402-1-AP) were purchased from Proteintech Group (Wuhan, Hubei, China). Antibodies against p-MLKL (ab196436), ADAMTs5 (ab41037) and MMP13 (ab39012) were obtained from Abcam (Cambridge, United Kingdom). Mouse SDF-1a/CXCL12 ELISA Kit was purchased from Bangyi (Shanghai, China).