Fig. 6. Validation of dependency of GBM cells on specialized protein kinases.
a, Viability curves of PDOs, each derived from an independent patient. Each curve represents one independent PDO assayed for the indicated compound or IR. Data in each curve are mean ± s.d. of n = 3 or 6 technical replicates for compound treatment (Source Data Fig. 6) and n = 8 technical replicates for IR. Experiments were performed twice with similar results. b, Viability curves of GPM PDOs (n = 14 PDOs, each derived from an independent patient) treated with BJE6-106. Data in each curve are mean ± s.d. of n = 6 or 18 technical replicates for each PDO (Source Data Fig. 6). The experiment was repeated three times with similar results. c, Colony-forming assay using GPM PDO cells treated with BJE6-106. Data are the mean of n = 3 technical replicates from one representative experiment. Experiment was repeated twice with similar results. CTRL, control. d, Western blot of GPM PDO cells treated with 50 μM of BJE6-106. Experiment was repeated twice with similar results. e, Western blot of GPM PDO cells transduced with lentivirus expressing two independent shRNAs targeting PRKCD or non-targeting shRNA (NT). Experiment was repeated three times with similar results. f,g, Growth curves of two independent GPM PDOs, PDO 019 (f) and PDO 008 (g) transduced as in e. Data are mean of n = 5 (f) and n = 6 (g) technical replicates from one representative experiment. Experiments were repeated twice with similar results. h, Quantification of sphere-forming assay for GPM PDO cells (PDO 008) transduced as in e. Data are mean ± s.d. of n = 3 independent infections/biological replicates. i, Rate of glucose uptake in GPM PDO cells (PDO 019) transduced as in e. Data are mean ± s.d. of n = 6 for shRNA NT, n = 3 for shPRKCD 1 and n = 4 for shPRKCD 2 technical replicates from two independent infections/biological replicates. j, Concentration of triacylglycerol in GPM PDO cells (PDO 019) transduced as in e. Data are mean ± s.d. of n = 4 for shRNA NT, n = 3 for shPRKCD 1 and n = 6 for shPRKCD 2 technical replicates from two independent infections/biological replicates. k, Cell viability after IR minus or plus nedisertib of PPR PDOs (n = 8 PDOs, each derived from an independent patient) and GPM PDOs (n = 8 PDOs, each derived from an independent patient). Data in each curve are mean of n = 4 technical replicates. Experiment was repeated twice with similar results. l, Western blot of PPR PDO cells treated with IR (4 Gy) or IR plus nedisertib (556 nM). Experiment was repeated twice with similar results. m, Quantification of γ-H2AX foci per nucleus in PPR PDO cells (PDO 044) after treatment as in l; the number (n) of nuclei is indicated (Source Data Fig. 6). Data are mean ± s.e.m. In each quantitative experiment, significance was established by two-tailed t-test, unequal variance or the Mann–Whitney test for experiment in m. In western blots, vinculin and β-actin are shown as loading controls.