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. 2016 Jul 27;25(18):3960–3974. doi: 10.1093/hmg/ddw237

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© The Author 2016. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — Induction of apoptosis in C2C12 myotubes induces indiscriminate ex-miRNA release. C2C12 cells were differentiated for 6 days and then apoptosis was induced by incubation with 1 µM Staurosporine (SP) for 24 h. (A) RNA was extracted from conditioned cell culture supernatants and miRNA levels were determined by RT-qPCR. Fold changes in miRNA abundance after SP treatment are shown relative to DMSO-treated control cultures. (B) Cell viability was determined by MTS assay and is reported as a percentage of the DMSO-treated control group. (C) Detection of apoptosis by Annexin V-FITC and Propodium Iodide (PI) fluorescence staining in Staurosporine treated (upper panel) and control cells (lower panel). Scale bar represents 200 µm (20x magnification). Apoptosis was further assessed using Trypan Blue staining to measure cell viability. All values are mean +SEM. *P<0.05, ***P<0.001 (unpaired t-test, 2 tailed, n=3).