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. 2017 Jul 10;7:4942. doi: 10.1038/s41598-017-05232-0

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© The Author(s) 2017

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Mitochondrial elongation after a 12 hr stimulation with lipopolysaccharide (LPS) is mediated by reactive oxygen species (ROS) and activated AMP-activated protein kinase (AMPK). Primary microglial cultures were untreated (Cont) or treated with LPS for 2 (2hr) or 12 (12hr) hr with or without drugs including a ROS scavenger (N-acetyl cysteine; NAC) and Compound C (Comp C), and then subjected to Western blotting and immunostaining for TOM20 (a). Western blotting of AMPK phosphorylated at Thr172 (p-AMPK) and total AMPK following LPS stimulation (b). Western blotting of p-AMPK with or without NAC treatment (c, left) and quantification of the western blotting analyses (c, right) are shown. *p < 0.05, **p < 0.01 in t-test (N = 6). Each dot represents each replicate and means (bars) are shown. Representative images (d–g) and the measured lengths (h) of TOM20-immunopositive mitochondrial profiles with or without treatments of NAC (h). The graphical summary of the model is shown (i). N = 1818 (control), 3489 (LPS 2hr), 2238 (LPS 12hr), (LPS 2hr + Comp C) and 3654 (LPS 12hr + Comp C) mitochondria in (h). ****p < 0.0001 in the U-test. Medians (h, bars) with quartile ranges (h, boxes) are shown. Bars: 20 μm.