Pbs 1x

Dimethyl sulfoxide (DMSO) and T-2 toxin from Fusarium sp. (cat. No T4887) were obtained from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Dulbecco’s modified Eagle medium (DMEM) with 4.5 g/L glucose and L-glutamine, heat-inactivated fetal bovine serum (FBS), penicillin–streptomycin mixture, and PBS (1X) without calcium or magnesium were purchased at Lonza (Basel, Switzerland). The 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazol-carbocyanine iodide (JC-1) probe, Hank’s Balanced Salt Solution (HBSS), 2′,7′-dichlorodihydrofluorescein diacetate (H₂DCFDA) probe and JC-1 dye were from Thermo Fisher Scientific (Waltham, MA, USA). All other chemicals were of molecular grade or the highest quality available.

Careful dissection was employed to isolate as much as possible of the cancer and the healthy tissue, both trimmed to have the same mass, which was assessed with mg accuracy (average sample mass 0.282±0.052 gr; range: 0.0432 gr–0.676 gr; n= 13). The samples were then carefully washed with phosphate buffered saline (PBS, 1X, Lonza) supplemented with penicillin (100 U/mL), streptomycin (100 μg/mL), and amphotericin B (250 μg/mL) at 1% (Lonza), to eliminate as much blood, feces residue, and other organic debris as possible, and to further sterilize the samples. The cleanliness of the latter was routinely checked by viewing them on a screen connected to a high-sensitivity digital CMOS camera, using a 2.3 megapixel CMOS sensor (C11440-36U, Hamamatsu Photonics, Tokyo, Japan) coupled to a microscope (TE 300, Nikon, Tokyo, Japan) with high magnification (Figure S1). Further details of sample handling are described in the supplementary materials.