Cyp7a1

Reagents were purchased as follows: DDC (Sigma-Aldrich, 137030), DMSO (Sigma-Aldrich, 543900), LPS (Sigma-Aldrich, L2630), ATP (YEASEN, 60605ES03), calcipotriol (MCE, HY-10001), calcifediol (MCE, HY-32351), MCC950 (MCE, HY-12815); The antibodies used for western blot were purchased as follows: NLRP3 (Cell Signaling Technology, 15101), caspase-1 p20 (Santa Cruz Biotechnology, sc-398715), ASC (Santa Cruz Biotechnology, sc-514414), IL-1β (Cell Signaling Technology, 12426), α-SMA (ABclonal, A17910), Col1A1 (ABclonal, A1352), β-actin (ABclonal, AC026), YAP1 (proteintech, 13584-1-AP), p-YAP1 (Cell Signaling Technology, 13008), VDR (ABclonal, A2194), CYP7A1 (ABclonal, A10615), MRP3 (Cell Signaling Technology, 39909), CYP8B1 (Abcam, ab191910). The antibodies used for IHC were purchased as follows: NLRP3 (ABclonal, A12694), α-SMA (ABclonal, A17910), Col1A1 (ABclonal, A1352).

Western blot was performed as previously reported [14 ]. The proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene fluoride (PVDF) membrane (Gendepot). The membrane was incubated following primary antibodies: p-AMP-activated protein kinase (AMPK), AMPK (Cell signaling, Beverly, MA, USA), sterol regulatory-element binding proteins (SREBP)1c (Santa Cruz Biotechnology, Dallas, TX, USA), fatty acid synthase (FAS; Cell signaling), peroxisome proliferator-activated receptor-γ (PPARγ; Cell signaling), CCAAT/enhancer-binding protein-α (C/EBPα; Cell signaling), SREBP1 (Abcam), SREBP2 (ABclonal, Wuhan, Hubei, China), cytochrome P450 family 7 subfamily a member 1 (CYP7A1; ABclonal), 3-hydroxy-3-methyl-glutaryl coenzyme A reductase (HMCGR; Santa Cruz Biotechnology), β-actin (Santa Cruz Biotechnology), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Santa Cruz Biotechnology). The secondary antibodies, including Goat anti-Mouse IgG (H+L) and Goat anti-Rabbit IgG(H+L)-HRP, were obtained from Gen-depot. The blots were visualized using the West-Q Femto Clean enhanced chemiluminescence (ECL) solution (Gendepot) and LuminoGraph III Lite (ATTO, Tokyo, Japan), following the manufacturer’s instructions. Quantitative analysis was measured with CS Analyzer 4 (ATTO).

Torularhodin (purity > 95%) was isolated and purified from the extract of Sporidiobolus pararoseus (JD-2 CCTCC M 2010326) according to a previously published method [10 (link)]. Animal diets were purchased from TROPHIC Animal Feed High-tech Co., Ltd. Commercial kits for serum concentration of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL-Cc), and triglyceride (TG) were purchased from Jiancheng Technology Co. (Nanjing, China). Hematoxylin and eosin (H&E) were obtained from Servicebio Technology Co. (Wuhan, China). Antibodies of PPARα, PPAR-γ, CYP7A1, CPT1A, SLC27A4, and GAPDH were obtained from ABclonal Technology Co. Ltd. (Wuhan, China). Western fluorescence detection reagent was obtained from Beyotime Biotechnology (Shanghai, China).