Nim811

GBM cell lines (U251 and T98G) were grown in DMEM, 10% FBS. G22VF cells were generated as previously described,⁴⁴ and were also grown in the same media as U251 cells. Reagents (cycloheximide, blasticidin, Qvd-oph, chloroquine, tunicamycin and thapsigargin) were purchased from Sigma-Aldrich (St. Louis, MO, USA). U0126 and bafilomycin-A1 were purchased from InvivoGen (San Diego, CA, USA). NIM811 was requested from Novartis (Basel, Switzerland). Rapamycin was from Alfa Aesar (Ward Hill, MA, USA), and torin-2 was obtained from Tocris Bioscience (Bristol, UK).

Mouse antibody against AIF and rabbit antibodies against p53, H2B, and Cyp40, respectively, were purchased from Santa Cruz Biotechnology; mouse antibody against CytC and rabbit antibodies against VDAC and CypA were from Abcam. Monoclonal mouse anti-E1 antibody was from Viral Antigens, and mouse anti-alpha tubulin was obtained from Sigma Aldrich (St. Louis, MO, USA). Secondary antibodies for immunofluorescence and Western blot analysis were from Dianova. The pan caspase inhibitor z-VAD-fmk was from PeptaNova (Sandhausen, Germany); PFTμ and α, staurosporine and camptothecin were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). NIM811 was generously provided by Novartis (Basel, Switzerland). All other reagents were from Sigma Aldrich. Stock solutions were prepared in dimethyl sulfoxide (DMSO), stored at −20 °C and diluted in cell culture medium to their respective final concentrations directly before use. Final concentration of DMSO, which was employed as vehicle (solvent) control, never exceeded 0.1%.