Z vad fmk

HEK-293T (ATCC Manassas, VA USA, CRL-11268), DLD-1 (ATCC, CCL-221), and SK-MEL-28 (ATCC, HTB-72) cells were maintained in Dulbecco’s Modified Eagle Media (DMEM) (Gibco/Thermo Fisher Scientific, Waltham, MA USA), supplemented with 10% fetal bovine serum (FBS), 100 μg/ml streptomycin and 100 units/ml penicillin (both from Merck, Darmstadt, Germany). HeLa cells (ATCC CCL-2) were maintained in Roswell Park Memorial Institute 1640 Media (RPMI) (Gibco), supplemented with 10% FBS, 100 μg/ml streptomycin, and 100 units/ml penicillin. The cells were incubated in cell culture flask at 37 °C with 5% CO₂. For experiments, cells were cultured in 6- or 12-well cell culture plates. To induce cell death, the following compounds were employed: A-Hemolysin from Staphylococcus aureus (α-toxin), Doxorubicin, Etoposide, Staurosporine, Nocodazol and Taxol (Merck, Darmstadt, Germany). In order to inhibit caspase activity, the following compounds were used: AQZ-1/AZ 10417808 (Tocris, Bristol, UK), z-VAD-fmk, and z-VDVAD-fmk (Biozol, Eching, Germany). For inhibition of p53 activity, the following compound was used: Pifithrin-α (Tocris, Bristol, UK).

TH34 (3-(N-benzylamino)-4-methylbenzhydroxamic acid) (stock concentration 50 mM) was synthesized by coauthors TH and WS as described previously (Heimburg et al. 2016 (link), 2017 (link)) and was dissolved in DMSO (Sigma-Aldrich). All-trans retinoic acid (ATRA, Sigma-Aldrich, stock concentration 10 mM) was dissolved in ethanol (EtOH, Sigma-Aldrich). Z-VAD-FMK (Biozol, Eching, Germany, stock concentration 100 mM), necrostatin-1 (Cayman Chemical, Tallinn, Estonia, stock concentration 50 mM) and trolox (kindly provided by N. Brady, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA, stock concentration 50 mM) were dissolved in DMSO. N-Acetylcysteine (NAC, Sigma-Aldrich, stock concentration 1 mM) was dissolved in autoclaved Millipore H₂O and stored at 4 °C protected from light. If not otherwise specified, compounds were stored at − 20 °C and protected from light.