A113 1

The levels of total triglyceride (TG), total cholesterol (TC), high-density-lipoprotein cholesterol (HDLC), low-density-lipoprotein cholesterol (LDLC), and total bile acid (TBA) in serum were measured by analysis kits (A110-2, A111-2, A112-1, A113-1, and E003-2-1, Jiancheng Institute of Biotechnology, Nanjing, China). Non-esterified fatty acid (NEFA) was measured by an automatic biochemical analyzer (Olympus Au5400). The concentrations of very-low-density-lipoprotein-cholesterol (VLDL-C) and lysophosphatidylcholine (LPC) were measured by enzyme-linked immunosorbent assay kits (H249, Jiancheng, Nanjing and CEK621Ge, Cloud-clone corp, Wuhan, China, respectively) according to the instruction manual.

After treatment with RA-XII, HepG2 cells were harvested, washed with PBS, and lysed in 1% Triton X-100 lysis buffer (Beyotime, Nanjing, Jiangsu, China) for 30 minutes. Tumor tissues were weighted and homogenized at 2500 rpm/min for 10 min, then centrifuged, and the supernatants were harvested. Protein concentration was assessed by BCA kit (Thermo, Rockford, IL, USA). TC, TG, LDL and HDL in the HepG2 cell lysates or solid tumors were assayed using commercial kits (A111-1, A110-1, A113-1, A112-1, Jiancheng, Nanjing, China) according to manufacturer’s specifications. This study set blank wells, standard wells, and detected sample wells. The blank wells were added with sample diluent and other wells were added with standard preparation or detected samples, respectively. For TC and TG detection, all wells were then filled with working solution at 37 °C for 10 min and Optical density (OD) values were measured at 510 nm. For LDL and HDL detection, all wells were then filled with working solution A and working solution B at 37 °C for 5 min successively and OD values were measured at 546 nm two times.