Fuji dri chem slide gpt alt piii

Wild‐type or Pml‐knockout C57BL/6 mice aged 6–8 weeks and of the same sex were used for TNF‐induced septic shock. Mice were anesthetized using Avertin, and mouse TNF (1.5 µg/g) in a total volume of 200 µl endotoxin‐free PBS was injected intravenously (i.v.). Body temperatures were monitored rectally every 1–3 h for 30 h using an industrial electronic thermometer (Kane‐May), and mouse mortality was recorded at the same time. Mice were sacrificed when their body temperature fell below 22°C. Serum was collected by cardiac puncture 15 min within mice dying. For live mice, serum was collected 30 h after TNF injection. Serum LDH was measured using an LDH Cytotoxicity Detection Kit (Clontech, #MK401) following the user manual. Serum GPT/ALT levels were determined using FUJI DRI‐CHEM SLIDE GPT/ALT‐P III (FUJIFILM, Tokyo, Japan). Mouse serum TNF, IL‐6, and IL‐1α levels were detected using a TNF alpha Mouse Uncoated ELISA Kit (Invitrogen, #88‐7324‐88), IL‐6 Mouse Uncoated ELISA Kit (Invitrogen, #88‐7064‐88), or ELISA MAX™ Deluxe Set Mouse IL‐1α (BioLegend, #433404), respectively. For MK2 and RIPK1 inhibitor treatment, mice were pre‐treated with 75 µg PF‐3644022 (Sigma‐Aldrich, # PZ0188) per mouse or Nec‐1s (6 µg/g) 15 min before TNF injection by intraperitoneal injection, and then subjected to inhibitor treatment again 60 min after TNF injection.