Alt c009 2 1

The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in mouse serum and cell supernatant were evaluated using the ALT (c009‐2‐1) and AST (c010‐2‐1) detection kits from NanJing JianCheng Bioengineering Institute (Nanjing, China) strictly in accordance with the protocol. The contents of TNF‐α and IL‐1β levels in serum and cell supernatant were examined using ELISA kits from Mskbio (Wuhan, China) (TNF‐α: 69‐99985; IL‐1β: 69‐59812). Briefly, the samples were centrifuged at 4000 rpm/min for 10 minutes at 4°C, followed by the collection of the supernatant. Then, the standard samples were added with 2 mL of distilled water to prepare a 20 ng/mL standard sample solution. For this purpose, 8 standard tubes were set, in which the first tube was added with a 900 μL diluted sample solution, whereas the rest of the tubes were added with a 500 μL sample solution. The content in each tube was repeatedly diluted with the eighth tube set as a blank control. Then, each well was added with 100 μL standard or test samples and placed on the reaction place at 37℃ for 120 minutes. Each sample was detected following the instructions of the ELISA kit. The corresponding IL‐9 content was determined on the curve based on the sample optical density (OD) value.