a, SU-pcGBM2 cells (EdU, red; DAPI, blue) exposed to conditioned medium generated in the presence or absence of ADAM10 inhibitor. Scale bar, 50 μm. b–d, Proliferation indices of SU-pcGBM2 cells exposed to plain medium (aCSF) or active conditioned medium generated in the presence or absence of pan-MMP inhibitor (BAT) (b), ADAM10 inhibitor (c) or ADAM10 inhibitor with or without NLGN3 rescue (d). n = 3 wells per condition. e, Cell viability of SU-pcGBM2 cells exposed to ADAM10 inhibitor (GI254023X, 10 nM–2 μM) at 24, 48 and 72 h (n = 3 wells per condition). f, Proliferation index of SU-pcGBM2 cells exposed to GI254023X (0–2 μM) (n = 3 wells per condition). g, Spheroid invasion index of SU-DIPG-VI cells exposed to ADAM10i (0–5 μM) at 24, 48 and 72 h expressed as the diameter of the sphere of glioma cells relative to the initial diameter at time 0 h. h, Neurosphere formation assay in SU-pcGBM2 cells in the presence of GI254023X (0–2 μM; n = 10 wells per condition. i, Extreme limiting dilution assay (ELDA) data presented in h re-plotted here as a log fraction plot with the slope of the solid line representing the log-active cell fraction and confidence intervals shown as dotted lines. SU-pcGBM2 cells treated with ADAM10 inhibitor GI254023X at 0.5 μM (black), 1 μM (red) or 2 μM (green), with vehicle (DMSO) control (royal blue) or no DMSO (cyan) and analysed for neurosphere formation at 2 weeks. In b–d and f, P values as indicated; one-way ANOVA with Tukey’s post hoc test for multiple comparisons. Data are mean ± s.e.m. In h, χ2 test; data are mean ± confidence intervals. n.s. indicates P > 0.05.