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. 2024 Jun 20;6(1):vdae104. doi: 10.1093/noajnl/vdae104

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© The Author(s) 2024. Published by Oxford University Press, the Society for Neuro-Oncology and the European Association of Neuro-Oncology.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.

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Figure 1. — Differential response of glioma cell lines to MLN4924 is independent of NEDD8 activating enzyme (NAE) targeted inhibition. (A) Viability of LN18, GB1, M059K, and SNU1105 cell lines 72 hours after MLN4924 treatment was measured. Data are normalized to DMSO (5 replicates). P < .001 by non-linear regression fit comparison. (B) Colony formation by LN18 and SNU1105 cell lines 2 weeks after MLN4924 treatment (24 hours; 5 replicates). Colonies (> 50 cells) were counted and reported as surviving fractions (mean ± SD, *** P < .001 by Student’s t-test). Drug dose–response assay of Bortezomib was conducted on glioma models (C) and TERT-immortalized normal human astrocytes (D). (E) Indicated cell lines were treated with increasing concentrations of MLN4924 for 8 hours; isolated proteins were subjected to Immunoblot (IB) using antibodies (Ab) against Nedd8-cullin or NAE1 of proteins from glioma cells after MLN4924 treatment (alpha-tubulin is loading control). (F) IB analysis of selected CRL substrates and DNA damage proteins from GB1 and M059K exposed to 500 nM MLN4924 for indicated times, (GAPDH is loading control).