Fetal calf serum (fcs)

RMS cell lines RH30, RD, RMS-YM, RH4, JR-1, RMS-01, RH41, HFF-1 have been described previously [17 (link)]. Cell lines were cultured in Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific, Waltham, MA, USA) (RD, RH30, RMS-01, RH4) or Roswell Park Memorial Institute 1640 (RPMI) medium (Thermo Fisher Scientific, Waltham, MA, USA)(RH41, RMS-YM) supplemented with 10% fetal calf serum (Gibco, Life Technologies Ltd., Thermo Fisher Scientific, Waltham, MA, USA), 2 mM L-glutamine (Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin/streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). HFF-1 cells were purchased from ATCC and cultured in Dulbecco’s modified Eagle’s medium supplemented with 15% fetal calf serum, 2 mM L-glutamine and 1% penicillin/streptomycin. Cells were maintained at 37 °C at 5% CO₂.
Cell lines were authenticated using the Geneprint 10 kit according to manufacturer’s instructions (Promega, Madison, WI, USA) and subsequent fragment length analysis was carried out by Eurofins Genomics (Ebersberg, Germany). The resulting short tandem repeat (STR) typing results were compared with public databases.

Tetracycline (Tet)-inducible HEK293-TREx cells, stably transfected with human TRPM7, were cultured in DMEM medium (Gibco) containing 10% fetal bovine serum (FBS, Gibco), zeocin (0.4 mg/ml, Invitrogen) and blasticidin (5 µg/ml, Invitrogen). Overexpression was induced adding 1 µg/ml Tetracycline (Gibco) to the culture medium. Patch-clamp experiments were performed 18–22 h post-Tetracycline induction. The normal rat kidney fibroblast cell line NRK-49F and epithelial cell line NRK-52E (ATCC) were maintained in DMEM medium (ATCC) supplemented with 5% fetal calf serum (ATCC). Cells were maintained at 37 °C under 5% CO₂ condition.

Dimethyl sulfoxide (DMSO), N-formyl-Met-Leu-Phe (fMLF), Trp-Lys-Tyr-Val-Met (WKYMVM), and Histopaque 1077 were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). (+)-Cedrol was from TargetMol (Boston, MA, USA). n-Hexane was purchased from Merck (Darmstadt, Germany). Fluo-4AM was purchased from Invitrogen (Carlsbad, CA, USA). Roswell Park Memorial Institute (RPMI) 1640 medium and Dulbecco’s Modified Eagle’s Medium (DMEM):F12 medium were purchased from HyClone Laboratories (Logan, UT, USA). Fetal calf serum and fetal bovine serum were purchased from ATCC (Manassas, VA, USA). Hanks’ balanced salt solution (HBSS; 0.137 M NaCl, 5.4 mM KCl, 0.25 mM Na₂HPO₄, 0.44 mM KH₂PO₄, 4.2 mM NaHCO₃, 5.56 mM glucose, and 10 mM HEPES, pH 7.4) was purchased from Life Technologies (Grand Island, NY, USA). HBSS without Ca²⁺ and Mg²⁺ is designated as HBSS^–; HBSS containing 1.3 mM CaCl₂ and 1.0 mM MgSO₄ is designated as HBSS⁺.

1205 Lu cells were maintained in melanoma medium, consisting of four parts of MCDB153 (Sigma, St. Louis, MO) and one part of L–15 (Life Technologies, Carlsbad, CA), supplemented with 2 mM CaCl₂, 5 μg/ml insulin (Sigma) and 2% fetal bovine serum (Invitrogen), 1% streptomycin/ampicillin and maintained at 37°C, 2% CO₂ under controlled humidity. 293T (ATCC^® CRL–11268™), T98G (ATCC^® CRL–1690™), and U87 MG (ATCC^® HTB–14™) cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal calf serum (FCS) at 37°C, 2% CO₂ under controlled humidity. Both U87 MG and T98G are well-established models for the study of GBM biology and molecular pathways [51 , 52 (link)]. U87 MG is a cell line established from a GBM sample from a 44 years old Caucasian male, highly invasive and able to produce tumors in nude mice [53 , 54 (link)]. On the other hand, T98G is a cell line established from a GBM sample from a 61 years old Caucasian male, the cells undergo G1 phase cell cycle arrest under stationary conditions and are unable to produce tumors in nude mice, but present anchorage-independent growth [55 (link)]. Human Normal Total RNA Master Panel II (Lot. No. 7090015) was purchased from Clontech (Palo Alto, CA).