Lx 2 cells

Mouse embryonic fibroblast cell line NIH3T3 cells were purchased from American Type Culture Collection. Human hepatic stellate cell line LX-2 cells were purchased from Merck Millipore. NIH3T3 cells, cardiac myofibroblasts, and activated HSCs were cultured at 37 °C with 5% CO₂ in Dulbecco’s Modified Eagle’s Medium (DMEM; Nacalai Tesque) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific) and 1% penicillin-streptomycin solution (Nacalai Tesque). LX-2 cells were cultured at 37 °C with 5% CO₂ in DMEM supplemented with 2% FBS and 1% penicillin-streptomycin solution.
Lipofectamine RNAiMAX (Thermo Fisher Scientific) was used for siRNA transfection in NIH3T3 cells, LX-2 cells, cardiac myofibroblasts, and activated HSCs, as per the manufacturer’s instructions. For drebrin overexpression in NIH3T3 cells, Lipofectamine 2000 (Thermo Fisher Scientific) was used as per the manufacturer’s instructions.

LX-2 cells (Merck Millipore, Burlington, USA), an immortalised human aHSC line [11 (link)], were maintained in Dulbecco’s modified eagle medium (DMEM, ThermoFisher Scientific, Waltham, USA) supplemented with 2% foetal bovine serum (FBS, Sigma-Aldrich, St. Louis, USA), 100 units/ml penicillin/streptomycin (ThermoFisher Scientific) and 4 mM L-glutamine (L-Glu, ThermoFisher Scientific) at 37 °C and 5% CO₂. Upon reaching ~ 80% confluency, LX-2 cells were detached from the culture flask using 0.25% trypsin-EDTA solution (ThermoFisher Scientific) and re-seeded according to a split ratio of 1:3.

The LX-2 cell line was obtained from Shanghai Institutes for Biological Sciences (Shanghai, China). The LX-2 cells were maintained in Dulbecco’s Modified Eagle Medium containing Nutrient Mixture F-12 (DMEM/F-12) (Gibco, Life technologies, New York, USA) supplemented with 10 % Fetal calf serum (Gibco, Life technologies, New York, USA). The cells were incubated at 37 °C at an atmosphere of 5 % CO₂. For stimulation, the cells were plated on 6 cm dishes starved in medium containing 1 % FCS. The cells were divided into two groups and for the control group, no treatment was performed. For the HCF group, stimulation with HCF for 48 h was carried out.
HCF was obtained by aspiration under aseptic conditions from liver cysts found in clinical CE patients with surgery. The hydatid fluid was centrifuged at 1000 g at 4 °C and the supernatant stored at −80 °C until use. The total protein concentration of HCF was determined using the commercially available bicinchoninic acid (BCA) assay (Thermo Fisher Scientific, Waltham, MA, USA).