Ht29 cell line

The HT29 cell line was obtained from LONZA (Basel, Switzerland). NCM460 cells were from INCELL Corporation (San Antonio, TX, USA). SW480 cells were a kind gift from Prof. Alberto Muñoz (CSIC, Madrid, Spain). Dulbecco’s modified Eagle’s medium (DMEM), penicillin, streptomycin, and fetal bovine serum (FBS) were sourced from Lonza (Basel, Switzerland). L-glutamine was from Gibco (Barcelona, Spain). Trypsin-EDTA was from LONZA (Verbiers, Belgium). Poly-L-Lysin was from Marlenfeld GmbH (Lauda-Könlgshofen, Germany). Six-well plates were from NUNC (Thermo Scientific, Waltham, MA, USA). Dishes 10 cm² in diameter were from Corning (NY, USA). DFMO was from TOCRIS (Bristol, UK). Fura2/AM and qPCR primers are from Invitrogen (Eugene, OR, USA). Cyclopiazonic acid (CPA) was from Sigma-Aldrich (Steinheim, Alemania). Antibodies against MCU and β actin were from Sigma (Madrid, Spain). The RNA extraction kit was a GeneMATRIX Universal RNA Purification Kit from EURx (Gdansk, Poland). Clariom D human microarrays (Affymetrix) were supplied by CABIMER (Andalucía, Spain). RNA-seq (Illumina) was provided by Sistemas Genómicos S.L (Valencia, Spain). PolyamineRED was from Funakoshi Co., Ltd., Tokyo, Japan). All other reagents were obtained from Sigma and Merck.

The HT-29 cell line from lonza (Verviers, Belgium) was consistently cultured in McCoy’s medium, enriched with 10% (v/v) heat-inactivated bovine fetal serum and a combination of antimicrobial agents (including 50 μg/mL penicillin, 50 μg/mL streptomycin, 50 μg/mL gentamicin, and 1.25 μg/mL amphotericin B). SigmaAldrich (St. Louis, MO, United States) provided all the necessary media and supplements. To maintain the cell line, standard procedures were followed: the culture medium was refreshed every 2 days, and the cells were trypsinized using a 0.25% trypsin–EDTA solution from SigmaAldrich (St. Louis, MO, United States) as needed.