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. 2021 Sep 1;22(17):9509. doi: 10.3390/ijms22179509

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Neddylation inhibition dysregulates SIRT1 intracellular localization. (A) Representative immunofluorescence images of Ctrl and NAE1-KO (KO1, KO2) C2C12 cells at 3 days in differentiation media. Cells were stained with SIRT1 (red), and nuclei were labeled with Hoechst 33342 (blue). Scale bar: 20 μm. (B) Western blots of whole cell lysates (WCL), cytosol, and nucleus fractions of Ctrl and NAE1-KO (KO1 and KO2) C2C12 cells at 3 days in differentiation media. GAPDH and LMNB1 were used as cytosol and nucleus marker proteins, respectively. (C) Quantifications of nuclear fraction of SIRT1 as normalized to LMNB1 with Ctrl set as 1. * p < 0.05 vs. Ctrl. One-way ANOVA. Three independent experiments.