60 mm culture plates

Caco-2 cells (ECACC, Salisbury, UK) were grown in MEM with Earle’s salts (PAA Laboratories GmbH, Pasching, Austria) containing 15% fetal bovine serum. HCT-116 cells (DSMZ, Braunschweig, Germany) were grown in McCoy’s 5A medium (Biowest, Nuaillé, France) containing 10% fetal bovine serum (PAA Laboratories). HCT-116 cells harbour an activating mutation in KRAS (G13D), while Caco-2 cells are KRAS wildtype. Culture media were supplemented with 2 mM glutamine, 1% non-essential amino acids, penicillin (100 U/ml), streptomycin (100 μg/ml) and amphotericin B (2.5 μg/ml). Cells were maintained in a humidified atmosphere at 37°C and 5% CO2. After cultures became 80% confluent (usually after 3 days), cells were trypsinized, centrifuged, and suspended in fresh medium. All cells used for experiments displayed > 95% viability and were seeded in 96-well plates, 6-well culture plates, 60-mm culture plates or ultra low-attachment plates (Corning Inc, Lowell, MA, USA). All experiments were carried out in duplicate and repeated at least three times.

HT-29 and HCT-116 cells (DSMZ, Braunschweig, Germany) were grown in McCoy's 5A medium (Biowest, Nuaillé, France) containing 10% fetal bovine serum (PAA Laboratories, Pasching, Austria). Caco-2 cells (ECACC, Salisbury, UK) were grown in MEM with Earle's salts (PAA Laboratories GmbH) containing 15% fetal bovine serum. DLD-1 cells (ATCC, US ) were grown in D-MEM high glucose medium (Capricorn scientific, GmbH), containing 15% fetal bovine serum. Culture media were supplemented with 2 mM glutamine, 1% non-essential amino acids, penicillin (100 U/ml), streptomycin (100 μg/ml) and amphotericin B (2.5 μg/ml) and cells were cultured in a humidified atmosphere at 37°C and 5% CO₂. in 96-well plates, 6-well culture plates, 60-mm culture plates or ultra low-attachment plates (Corning Inc, Lowell, MA, USA). All experiments were carried out in duplicate and repeated at least three times.