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MEC1 cells were provided by Dr. M. Hallek (Uniklinik Koeln). The immunophenotype matched the original description and the complex karyotype was highly similar to what was previously described [14 (link)]. p53 dysfunction was confirmed by the absence of p53 protein (Western blot) and absence of up-regulation of PUMA mRNA (RT-MLPA) and CD95 (FAS; fluorescence activated cell sorting [FACS]-staining) upon radiation (Supplementary Figure 2 to be found online at http://informahealthcare.com/doi/abs/10.3109/10428194.2014.996751). Cells were cultured in Iscove modified Dulbecco medium (IMDM; Gibco Life Technology, Paisley, UK) supplemented with 10% (vol/vol) heat inactivated fetal calf serum (FCS; ICN Biomedicals, Meckenheim, Germany), 100 μg/mL gentamycin and 5 mM L-glutamine (Invitrogen, Carlsbad, CA) at 37°C.
For CD40 stimulation, the CD40-ligand transfected cell line HeLa-CD154 (for control: mock-transfected HeLa cells) was used (American Type Culture Collection [ATCC], Manassas, VA) as described previously [15 (link)]. In the synergy experiments the following drugs and reagents were used: CDDP (Mayne Pharma, Brussels, Belgium), CpG oligonucleotide type B-Human TLR9 ligand (ODN2006; Invivogen, San Diego, CA), F-ara-A (Sigma-Aldrich, St Louis, MO), imatinib (Novartis, Basel, Switzerland) and CH11 (Beckham Coulter Company, Marseille, France).