Tissue total cholesterol assay kit

Liver tissues were washed three times with cold phosphate-buffered saline (PBS) and subjected to extraction with organic solvents (7:11:0.1, chloroform/isopropanol/Triton X-100). The TCHO and TBA were measured using the Tissue Total Cholesterol Assay Kit (E1015, Applygen, Beijing, China) and the total Bile Acid Assay Kit (STA-631, Cell Biolabs, Inc., CA, USA), and normalized to total protein concentrations.

The serum hormone of TC, TG, HDL-C, LDL-C, ALT, AST were measured with Hitachi 7100 automatic biochemical analyzer. And liver TC and TG levels were measured utilizing Tissue total cholesterol assay kit (Applygen Technologies Co. Ltd., China) and Tissue triglyceride assay kit (Applygen Technologies Co. Ltd., China), respectively.

The human umbilical vein endothelial cells (HUVEC, CRL-1730), human brain microvascular endothelial cells (HBMEC, CRL-3245), human aorta-vascular smooth muscle cells (HA-VSMC, CRL-1999), and human monocytes THP-1 (TIB-202) cell lines were purchased from the American Type Culture Collection (Manassas VA, USA); The oil red O solution was obtained from Beyotime Biotechnology (Shanghai, China); the shRNA lentivirus vector and the polybrene were obtained from the Cyagen Bioscience Inc., (Suzhou, China); IL-12, IL-6, and TNF-α ELISA kits were provided by Boster Biological Technology Co. Ltd. (Wuhan, China); Tissue Total cholesterol Assay Kit was from Applygen Technologies Inc. Beijing, China); PrimeScript RT reagent Kit, SYBR Premix DimerEraser™ (Perfect Real Time) assay kit, and the primers were purchased from Takara (Dalian, China); the primary antibodies p-TNK1 (D46E7), TNK1 (C44F9), p-STAT1 (58D6), STAT1 (D1K9Y), p-Tyk2 (Y1054), and Tyk2 (9312S) were purchased from CST (Danvers, MA, USA); the primary antibody for β-actin (AP0060) was purchased from Bioworld Technology (Nanjing, China); and the oxLDL was provided by the Peking Union-Biology, Co. Ltd (Beijing, China).

At age 12–13 weeks, eight female and male offspring in each group were killed. Blood was drawn from the inner canthus after the mice were fasted for 8 h. The serum was immediately separated and stored at −80°C for subsequent analysis or hormone determinations. Leptin and fasting insulin levels were measured with radioimmunoassay kits (Beijing North Institute of Biological Technology, Beijing, China). Lipid profiles were determined using biochemical analysis kits (China Diagnostics Medical Corporation, Beijing, China). After blood sample collection, each liver was immediately removed and stored at −80°C. Total cholesterol (CHO) and triglycerides (TG) were extracted from liver and measured with a Tissue Total Cholesterol Assay Kit and Tissue Triglyceride Assay Kit (Applygen Technologies, Beijing, China).