Ant060

After the cells were treated, they were lysed with RIPA (AntGene, ANT060) lysis buffer to extract the total protein in the cells. Then, the protein concentration was determined by the BCA protein detection kit, and a 10% SDS-PAGE gel was prepared for electrophoresis. After electrophoresis was completed, the proteins were transferred to a polyvinylidene fluoride membrane. A 5% skimmed milk solution was prepared and it was sealed with milk for 2 hours after the transfer was completed. Anti-LOX-1 (Abcam, ab203246), anti-MCP-1(NOVUS, NBP2-22115), anti-VCAM-1(GeneTex, GTX110684), anti-ZO-1(Cell Signalling Technology,13663S), anti-VE-cadherin (Cell Signalling Technology, 2500S), and anti-GAPDH (AntGene, ANT012) antibodies were incubated overnight at 4°C. A rabbit HRP conjugate was used as the secondary antibody. The optical density of the Western blot was quantified by using Image Lab software.

The expression of LOX-1 (Abcam, ab203246), ABCG5 (Boster, PB0381), ABCG8 (Boster, A01482) and SR-BI (GeneTex, GTX113645) in liver tissues were detected using western blot. Liver tissues were homogenized in RIPA buffer (AntGene, ANT060) supplemented with protease and phosphatase inhibitors at 4 °C for 30 min. After centrifugation at 12,000×g for 15 min at 4 °C, the supernatant was collected and measured using the PierceTM BCA Protein Assay Kit according to the instructions. After the system was prepared, it was electrophoresed on a 10% polyacrylamide SDS gel, and then transferred to the membrane. The membrane was blocked with 5% skimmed milk powder at room temperature for 2 h, and then it was incubated in the indicated antibody at 4 °C overnight. The next day, incubate with horseradish peroxidase-conjugated secondary antibody for 2 h at room temperature, and then observe using ECL Kit. Chemiluminescence was detected by exposure to film and quantified using Image Lab.