Penicillin mixture

Naringin (≥98%) was obtained from Baoji Herbest Bio-Tech Co., Ltd. (Baoji, China). THP-1 and RAW264.7 were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China). RPMI 1640 medium, fetal bovine serum (FBS), penicillin mixture and 0.25% Trypsin-EDTA were purchased from Gibco (Carlsbad, CA, USA). 2-Acetoxy-1-methoxypropane (PMA) and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). A cell counting kit-8 (CCK-8) was purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China). ox-LDL and Dil-ox-LDL were purchased from Yiyuan Biotechnology (Guangzhou, China). A free cholesterol (FC) content assay kit and a total cholesterol (TC) content assay kit were obtained from Solarbio Technology Co., Ltd. (Beijing, China). IL-1β, IL-6, IL-10, TNF-α and MCP-1 ELISA kits were obtained from BOSTER Biological Technology Co., Ltd. (Wuhan, China). A cholesterol efflux assay kit was purchased from Abcam (Cambridge, UK). Antibodies were purchased from ABclonal Technology Co., Ltd. (Wuhan, China).

The GCs were harvested from pre-hierarchical (6–10 mm), F4–F2, and F1 follicles. The culture medium used in the study consisted of DMEM/F12, 10% FBS (Gibco), 1% streptomycin, and penicillin mixture (Gibco). The cells resuspended with above medium were seeded on different culture plates and then incubated at 37°C under 5% CO₂ in humidified air to allow the cells to reach a confluence. After incubation of around 6 h, the medium was replaced by fresh medium. The six-well plates (∼15 × 10⁵ cells/well) were used for cell immunofluorescence assay, and the 12-well plates (∼10 × 10⁵ cells/well) were used for qPCR. In addition, the 96-well plates (∼5 × 10⁵ cells/well) were used for the collection of images and the growth curve of GCs. For three groups of GCs cultured in vitro, each experiment was conducted in triplicate.

The glioma cell line U251 was purchased from ATCC, USA. Fetal bovine serum, high glucose DMEM, penicillin mixture, and trypsin were all purchased from Gibco, USA. LCA powder (molecular formula: C₂₁H₂₂O₄; molecular weight: 338.40; purity ≥ 96.0%), dimethyl sulfoxide preparation, and methyl thiazolyl tetrazolium (MTT) kit were purchased from Sigma, USA. A 4% paraformaldehyde was purchased from Invitrogen, USA. Radio immunoprecipitation assay (RIPA) cell lysate was purchased from Beyotime Institute of Biotechnology, China. Bicinchoninic acid (BCA) kit and horseradish peroxidase-labeled goat anti-mouse IgG antibody were purchased from Thermo Company, USA. The ultrasensitive ECL chemiluminescence kit was purchased from Advansta, USA. Mouse anti-human Ki-67, β-catenin, GSK3β, Axin2, c-myc, Bcl-2, Bax, and GAPDH were all purchased from Cell Signaling, USA. Puromycin was purchased from Merck, USA. Lentiviral Vector Particle was purchased from China Jikai Gene. The cell culture dishes were purchased from Corning Costar, USA.