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. 2021 Oct;7(5):a006109. doi: 10.1101/mcs.a006109

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© 2021 Keddy et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed under the terms of the Creative Commons Attribution-NonCommercial License, which permits reuse and redistribution, except for commercial purposes, provided that the original author and source are credited.

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Figure 2. — Genetic analysis of tumors and in vitro characterization of identified NTRK3 mutations. (A) Variant allele frequency of identified mutations from GeneTrails next-generation sequencing of diagnostic and autopsy samples. (B) Immunoblot analysis of phosphorylated NTRK3, flag-tagged NTRK3, and phosphorylated and total ERK from transfected HEK293T17 cell lysates after 4 h of treatment with TKIs as indicated. (C) Bar graph of cell proliferation IC₅₀ values for each of the indicated TKIs against Ba/F3 cells expressing wild-type ETV6-NTRK3 (green) and ETV6-NTRK3^G623E (blue). (D) Scatter plot of cell proliferation IC₅₀ values for indicated TKIs against patient cell line BTO-4039 expressing ETV6-NTRK3^G623A cultured in 10% FBS media or neural stem cell media supplemented with bFGF and EGF. (E) Immunoblot analysis of phosphorylated NTRK3 and total NTRK3 from patient cell line BTO-4039 after 5 h of treatment with TKIs as indicated. (F) Predicted binding poses for entrectinib with NTRK3^WT, NTRK3^G623A, and NTRK3^G623E.