Lx 2 cells

hESC was generated in Dr. Jinsong Li’s laboratory, and cultured as previously described [47 (link)]. Human hepatic stellate cells (LX2 cells) was purchased from Procell (Life Science & Technology Co,. Ltd) in Wuhan, China, and cultured maintained as described in protocol from the provider.

The 98% purity of Schisandrin B was purchased from Shanghai Yuanye Biotechnology Co., Ltd (Shanghai, China). The Rat HSC line HSC-T6 and the human HSC line LX-2 cells were obtained from Procell Life Science and Technology Co.,Ltd (Wuhan, China). DMSO was purchased from Sigma-Aldrich (Shanghai, China) Trading Co, Ltd. DMEM, FBS and Penicillin Streptomycin were from Gibco/Invitrogen (Grand Island, NY, USA). Cell Counting Kit-8 were obtained from DOJINDO (Kumamoto Prefecture, Kyushu Island, Japan). TGF-β1 was purchased from Peprotech (Rocky Hill, NJ, USA). Anti-Caspase-3 (sc7272) antibody were purchased from Santa Cruz Biotechnology Inc (Santa Cruz, CA, USA). The anti-α-SMA (ab5694), anti-Collagen I (ab34710), anti-TIMP1 (ab61224), anti-BCL-2 (ab182858), anti-Bax (ab32563) anti-GAPDH (ab8245) antibody, horseradish-peroxidase (HRP) conjugated secondary antibody (a11008) and Fluoroshield Mounting Medium with DAPI were purchased from Abcam (Cambridge, MA, USA). The BCA Protein Assay Kit and BeyoECL plus kit was obtained from Beyotime (Shanghai, China). Goat anti-Rabbit IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 were obtained from ThermoFisher Scientific (Waltham, MA, USA). FITC/PI cell apoptosis kit were purchased from Keygen Biotechnology Co., Ltd (Nanjing, China).

Considering the ability of the LX-2 human HSC line to retain key features of hepatic stellate cytokine signalling and fibrogenesis, LX-2 cells (33 (link)) were purchased (Procell Life Sciences, China) and cultured in Dulbecco’s modified eagle medium with 10% foetal bovine serum (GIBCO, US) containing 1% penicillin and 1% streptomycin (GIBCO). DMEM media with and without glutamine was purchased from Procell Life Sciences. Before stimulation treatment, a cells were seeded at a density of 2×10⁵/mL on a six-well plate, and cultivate in serum-free medium for 8 hours, and hypoxic treatment consisted of 200 μM palmitic acid (PA, P0500, Sigma-Aldrich) in 1% O₂ for 72 h. Cell culture was conducted in CO₂ incubator (Heracell, Thermo, US) at 37°C, 5% of CO₂, and 21% of O_2; and in tri-gas CO₂ incubators (Memmert, Shanghai, China) at 37°C, 5% of CO₂, and 1% of O₂.

LX-2 cells were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China), and cultured in DMEM medium with 10% fetal bovine serum, 1% streptomycin, and 1% penicillin. The cells were placed in an MCO-15AC incubator (Sanyo, Tokyo, Japan) with culture parameters of 37 °C, 5% CO₂, and 95% air. Two days before the cell imaging experiment, we digested the cells with trypsin and put an appropriate amount of cells into the culture dish for the convenience of the subsequent experimental operation.