Mitopq

The patient-derived GBM cultures were obtained from the HGCC resource of Uppsala University, Sweden, which authenticates all primary cultures [25 (link)]. U3005/3013/3024/3028/3031/3034/3062MG passages 15−30 were cultured in N2B27 media (Thermo Fischer Scientific, Uppsala, Sweden) [Dulbecco’s Modified Eagle’s Medium (DMEM)/F12 Glutamax and Neurobasal medium [53 (link)] mixed at 1:1 ratio, with the addition of 1% B27 and 1% N2, 100 U/ml penicillin and 100 mg/ml streptomycin (Sigma-Aldrich Sweden AB, Stockholm, Sweden), 10 ng/ml EGF and 10 ng/ml FGF2 (PeproTech, EC Ltd, London, UK)]. Adherent cultures were seeded onto poly-ornithine/laminin-coated dishes and passaged as described [54 (link)]. Cells were treated with 1 µM aPKCi, (CRT-0066854-hydrochloride; TOCRIS/Bio-Techne Ltd, Abingdon, UK), 100 µM MitoPQ (mitochondria-targeted redox cycler; Abcam, Cambridge, UK) or their vehicle, dimethyl-sulfoxide (DMSO), and with 50 µM MitoTEMPO (SML0737; Sigma-Aldrich) or its vehicle, H₂O, as indicated below and in the figures

Deferoxamine (DFO), CoCl₂, 2-methoxyestradiol (2-ME), N-acetyl-L-cysteine (NAC), MitoTEMPO, U0126, 5-aza-2’-deoxycytidine (5-aza-dC), Y294002, SB203580, Ro31-8220, rapamycin, BML-275, trichostatin A (TSA) and 2’,7’-dichlorofluorescein diacetate (DCF-DA) were obtained from Sigma-Aldrich (St Louis, MO. USA), and MitoPQ, MitoSOX-Red and 2‐(N‐(7‐nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl)amino)‐2‐deoxyglucose (2‐NBD‐glucose) were from Abcam (Cambridge, UK), Thermo Fisher Scientific (Waltham, MA, USA) and Life Technologies (Carlsbad, CA, USA), respectively. AR-C155858 and SP600125 were supplied by Tocris Bioscience (Bristol, UK), and, syrosingopine was obtained from Extrasynthese SA (Lyon, France), respectively. TX-402 was a gift from Dr. H. Nagasawa, Gifu Pharmaceutical University. rapamycin was purchased from LC Laboratories (Woburn, MA, USA); BAY11-7082 was from AdipoGen Life Sciences (San Diego, CA, USA); PF-4708671 and KU-0063794 were from Selleck Chemicals LLC (Houston, TX, USA), and chaetocin was from Enzo Life Science (Farmingdale, NY, USA).