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. 2023 Nov 1;623(7986):366–374. doi: 10.1038/s41586-023-06678-1

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 8 — a, Western blot of proteins from SU-DIPG-VI cells treated with BDNF recombinant protein (100 nM) over a time course and probed for the indicated antibodies to demonstrate activation of downstream signaling pathways in comparison to untreated cells (vehicle only). b, Quantification of MAPK pathway activation in a, as the ratio of the normalized phospho-ERK (T202/Y204) levels to corresponding total protein levels for BDNF treated (100 nM) cultures compared to control (y-axis is in arbitrary units, n = 3 independent biological replicates, pERK Control vs 10 m P = 0.0048, Control vs 30 m P = 0.0094, Control vs 1 h P = 0.0167). c, Quantification of PI3K pathway activation in a, by comparing the ratio of the normalized phospho-AKT (S473) to corresponding total protein levels for BDNF treated (100 nM) cultures compared to control (y-axis is in arbitrary units, n = 3 independent biological replicates, pAKT Control vs 10 m P = 0.0293, Control vs 30 m P = 0.0307, Control vs 1 h P = 0.0148). d, Quantification of calcium pathway activation in a, by comparing the ratio of the normalized phospho-CAMKII (T286) to corresponding total protein levels for BDNF treated (100 nM) compared to control (y axis is in arbitrary units, n = 3 independent biological replicates, pCAMKII Control vs 10 m P = 0.0197, Control vs 30 m P = 0.0310, Control vs 1 h P = 0.0374). e, Representative Western blot analysis of primary patient-derived glioma culture, SU-DIPG-VI, treated with 100 nM BDNF at several timepoints using indicated antibodies. f, Quantification of the phospho-immunoblots ratio to corresponding total protein levels and normalized to vehicle treated control (y axis is in arbitrary units, n = 3 independent biological replicates, pGluA4 Control vs 10 m P = 0.0174, Control vs 30 m P = 0.0072), 8 h (pGluA4 Control vs BDNF P = 0.078). g, Representative Western blot analysis of 100 nM BDNF treated glioma cells (as in e,) at 30 min with and without entrectinib treatment (Ent, 5 µM). h, quantification of phospho-immunoblots (y axis is in arbitrary units, n = 3 independent biological replicates). Data are mean ± s.e.m. *P < 0.05, **P < 0.01, ns = not significant. Two-tailed one Sample t test for b, c, d, f, h.