Patient-Derived Glioblastoma Cell Culture

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