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p. administration of 700?mg/kg body weight of D-GalN and then treated with i.p. administration of 15?��g/kg body weight of hTNF��. All reagents were balanced with sterile PBS so that the total volume of injecting solution in different Roxadustat mice was the same. Mortality rate was recorded every 2?hr for up to 25?hr after the treatment. The animal protocols were approved by University of Chicago Institutional Animal Care and Use Committee. To analyze liver injury, livers were isolated from premoved mice, and the liver lobes were excised and fixed in 4% paraformaldehyde for 12?hr. The tissues were sliced to 5?��m thickness. H&E staining was performed at the University of Chicago Human Tissue Resource Center (HTRC). In situ cell death was analyzed by TUNEL staining (TUNEL Apoptosis Detection kit, EMD Millipore) according to the manufacturer��s protocol. The percentage of apoptotic cells among groups was determined Adenylate cyclase using Student��s t test. Mouse survival curves were constructed using the Kaplan-Meier product limit estimator and compared using the log rank (Mantel-Cox) test. p?Androgen Receptor Antagonist and apoptotic cells were analyzed by using FACS flow cytometer (BD Biosciences). Cytosol and mitochondria fractionations were prepared by the cytosol/mitochondria fractionation kit (Bio vision), according to the manufacturer��s instruction. NF-��B luciferase (LUC) reporter activity assays were determined as previously reported (Purcell et?al., 2001). Total RNA was used for quantitative real-time PCR, using the Absolute qPCR ROX Mix (Thermo Scientific) and Taqman PCR program. The PCR primers for detecting mRNA of IL-6 and I��B�� were synthesized from IDT. GAPDH was used as internal control. The primers were designed as the following: IL-6: sense, 5��-CAAAGCCAGAGTCCTTCAGAG-3��; antisense, 5��-GTCCTTAGCCACTCCTTCTG-3��; Probe, 5��- CCTACCCCAATTTCCAATGCTCTCCT-3�� I��B��: sense, 5��-AGGAGTACGAGCAAATGGTG-3��; antisense, 5��-CGGCTTCTCTTCGTGGATG-3��; Probe, 5��- CGGAGACTCGTTCCTGCACTTGG ?3��; GAPDH: sense, 5��-CTTTGTCAAGCTCATTTCCTGG-3��; antisense, 5��- TCTTGCTCAGTGTCCTTGC-3��; Probe, 5��- CACCCTGTTGCTGTAGCCGTATTCA-3��. We are grateful to Drs. David A. Brenner, Joseph A. DiDonato, Michael Karin, Stanley Korsmeyer, and Frank Mercurio for reagents that make this work possible.