Dmem glutamax l

NB cell lines were obtained from American Type Culture Collection (ATCC) (Manassas, VA, USA) and maintained in DMEM GlutaMAX^™-l or RPMI medium (Life Technologies, Inc., Darmstadt, Germany), supplemented with 10% fetal calf serum (FCS), 1% penicillin/streptomycin, 1 mM sodium pyruvate and 10 mM HEPES (all from Life Technologies, Inc.). LAN-5 cells were grown in flasks coated with rat tail I collagen (BD Biosciences, San Jose, CA, USA). Doxorubicin, VCR, VBL and VNR were purchased from Sigma (Deisenhofen, Germany); zVAD.fmk from Bachem (Heidelberg, Germany), Enbrel from Pfizer (Berlin, Germany), Nec-1 from Biomol (Biomol GmbH, Hamburg, Germany) and TNFα from Biochrom (Biochrom GmbH, Berlin, Germany). The bivalent Smac mimetic BV6 was kindly provided by Genentech (South San Francisco, CA, USA), chemicals were purchased from Sigma unless otherwise indicated.

NB cell lines with different MYCN status (SH-EP: MYCN non-amplified; LAN-5: MYCN amplified; NLF: MYCN amplified) were obtained from American Type Culture Collection (ATCC) (Manassas, VA, USA) and maintained in DMEM GlutaMAX™-l or RPMI medium (Life Technologies, Inc., Darmstadt, Germany), supplemented with 10% fetal calf serum (FCS), 1% penicillin/streptomycin, 1 mM sodium pyruvate and 10 mM HEPES (all from Life Technologies, Inc.). LAN-5 cells were grown in flasks coated with collagen (BD Biosciences, San Jose, CA, USA). VCR, VBL and VNR were purchased from Sigma (Deisenhofen, Germany); RO-3306 from Merck (Darmstadt, Germany); zVAD.fmk from Bachem (Heidelberg, Germany). MCL-1 plasmids (pCMV-Tag 3B, MCL-1 ‘4A’) were kindly provided by Genentech, Inc. (South San Francisco, CA, USA). Chemicals were purchased from Sigma unless otherwise indicated.

ES cell lines were kindly provided by C. Roessig (Muenster, Germany) or obtained from German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany) or American Type Culture Collection (Manassas, VA, USA) and authenticated by STR profiles. Cells were maintained in DMEM GlutaMAX^™-l or RPMI 1640 GlutaMAX^™-l medium (Life Technologies, Inc., Darmstadt, Germany), supplemented with 10% fetal calf serum (FCS), 1% penicillin/streptomycin and 1 mM sodium pyruvate. Cell lines were regularly tested for mycoplasma contamination to guarantee that experiments were performed only with mycoplasma-free cells. For colony formation assay, cells were seeded on collagen-coated plates. Eribulin was obtained from Eisai Inc. (Frankfurt, Germany), zVAD.fmk from Bachem (Heidelberg, Germany) and bortezomib from Selleckchem (Munich, Germany). PLK1 inhibitor BI 6727 was kindly provided by Boehringer Ingelheim (Vienna, Austria) and TRAIL receptor-2 agonistic antibody ETR2 from Human Genome Sciences (Rockville, MD, USA). Chemicals were purchased from Sigma-Aldrich or Carl Roth (Karlsruhe, Germany) unless otherwise indicated.