Hepg2

RIL175 WT cells were kindly provided by Prof. Simon Rothenfußer (Division of Clinical Pharmacology, University Hospital, LMU Munich, Germany) and had originally been isolated from hepatic tumors established in C57BL/6 mice as described previously (Müller et al., 2021 (link)). HUH7 cells were obtained from Japanese Collection of Research Bioresources (JCRB0403, Tokyo, Japan). HepG2 and Hep3B cells, and HUVECs were purchased from American Type Culture Collection (HB-8065, HB-8064, PCS-100-010 Manassas, Virginia, USA). Cancer cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM; RIL175 WT and TRPML1 KO, HUH7, HepG2) or Eagle's minimum essential medium (EMEM) (Hep3B), supplemented with 10% fetal calf serum (FCS) and 100 U/ml penicillin-streptomycin (Pan Biotech, Aidenbach, Germany). The cells were maintained at 37°C in a humidified atmosphere of 5% CO₂. None of the cell lines used are listed in the database of commonly misidentified cell lines maintained by ICLAC. All cells are proven to be mycoplasma-free on a quarterly basis.

We utilized seven cell lines in this study. The cell lines K562, SK-hep-1, HepG2, and CCD18co were purchased from the American Type Culture Collection (ATCC, Manassas, Virginia, USA). Whereas the cell lines Jurkat, U266 and OPM2 were acquired from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). We cultured K562, U266, Jurkat, and OPM2 in RPMI1640 medium (Pan-Biotech, Aidenbach, Bavaria, Germany) supplemented with 10% FBS (Sigma-Aldrich Chemie GmbH, Munich, Germany) and 1% penicillin/streptomycin (P/S) (Gibco, Schwerte, Germany). While SK-hep-1, HepG2, and CCD18co cells were maintained in EMEM medium (Pan-Biotech, Aidenbach, Bavaria, Germany) supplemented with 10% FBS (Sigma-Aldrich Chemie GmbH, Munich, Germany) and 1% penicillin/streptomycin (P/S) (Gibco, Schwerte, Germany). Meticrane (Sigma-Aldrich Chemie GmbH, Munich, Germany) was dissolved in DMSO and stored at -20°C at a concentration of 200mM. The HDAC inhibitor CUDC-101 (Selleck Chemicals GmbH, Munich, Germany) and the selective HDAC6 inhibitor ACY1215 (Cayman Chemical, Ann Arbor, Michigan, US) was dissolved in DMSO and stored at -20°C at a concentration of 50mM. Also, DNMT1 inhibitor 5-Azacytidine (5AC) (STEMCELL Technologies Germany GmbH, Cologne, Germany) was dissolved in DMSO and stored at -20°C at a concentration of 25mM.