Hcf cells

HCF cells were purchased from ScienCell Research Laboratories (San Diego, CA, USA) and cultured in Fibroblast Medium (ScienCell Research Laboratories) with Fibroblast Growth Supplement to ascertain the cell phenotypes during passages. Cells from passages 3–6 were used for the experiments. The cells were treated with 44 μM ATX for 6 h with or without pretreatment for 30 min with 1 µM HA130, an ATX inhibitor (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Primary human ventricular cardiac fibroblast (HCF) cells were purchased from ScienCell Research Laboratories; primary cells isolated from two individual donors were included in this study. Cells were cultured and maintained in Dulbecco modified Eagle medium (DMEM; Gibco, Leicestershire, UK), supplemented with 10% fetal bovine serum (Gibco, Leicestershire, UK) and penicillin–streptomycin antibiotics (Gibco) in a 5% CO₂ humidified incubator at 37 °C. Cells were not expanded beyond 15 population doublings in line with suggested conditions provided by ScienCell Research Laboratories. When required, HCF cells were treated for 72 h with either vehicle or 5 ng/mL human recombinant transforming growth factor β1 (TGF-β1; Promokine, Heidelberg, Germany) in serum free media. HCF cells were also stimulated with TGF-β1 in the presence of either 25 nM siRNA (siRNA) or 25 nM scrambled non-targeting control (scr), for genes that were identified as upregulated in DCM (LTBP2 and AEBP1).