*2.2. Cell Culture and Treatment*

The human monocytic cell line THP1-Lucia™ NF-κB, deriving from the human THP-1 monocyte cell line by stable integration of an NF-κB-inducible luciferase reporter construct, was purchased from Invivogen (San Diego, CA, USA) and HT-29, a human colorectal adenocarcinoma cell line from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). THP1-Lucia™ were cultured in RPMI 1640 medium, HT-29 in Dulbecco's Modified Eagle's Medium (DMEM), both supplemented with 10% (*v*/*v*) heat-inactivated fetal bovine serum and 1% (*v*/*v*) penicillin–streptomycin (100 U/mL). THP1-Lucia™ were treated alternately with zeocin and normocin (100 μg/mL; Invivogen, San Diego, CA, USA). Noncancer human colon epithelial cells HCEC-1CT [33,34] were kindly provided by Prof. Jerry W. Shay (UT Southwestern Medical Center, Dallas, TX, USA). HCEC-1CT cells were cultivated in high glucose DMEM. Basal medium was supplemented with the following components: Medium 199 (10<sup>×</sup>; 2% (*v*/*v*)), HyClone™ Cosmic Calf™ Serum (2% (*v*/*v*)), gentamicin (50 μg/mL), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (20 mM), insulin-transferrin-selenium-G (10 μg/mL; 5.5 μg/mL; 6.7 ng/mL), hydrocortisone (1 μg/mL) and recombinant human epidermal growth factor (18.7 ng/mL). All three cell lines were subcultured every 3–4 d, maintained in humidified incubators at 37 ◦C and 5% CO2 and routinely tested for the absence of mycoplasm contamination. Cell culture media, supplements and material were purchased from GIBCO Invitrogen (Karlsruhe, Germany), Sigma-Aldrich (Munich, Germany) and Sarstedt AG

& Co. (Nuembrecht, Germany). DON, NX-3, AURO and their combinations were added to the incubation solutions, resulting in a final solvent concentration of 1% (*v*/*v*) DMSO. In order to ensure data comparability, combinatory effects were always assessed on the same culture plate, in parallel to the individual substance.
