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. 2016 Sep 16;6:33647. doi: 10.1038/srep33647

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Copyright © 2016, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Spermatozoa were incubated in presence of bafilomycin A1 for 2 hours. Then, cells were prepared as described in material and methods. Negative controls are representative images from the head and the middle piece where primary Ab was omitted. Figures (a,b) are representative areas of the head. Figures (c,d,e) are digitally augmented images from mitochondria of the middle piece of the spermatozoa. Some of the LC3 proteins labeled with immunogold particles are marked with arrows; red arrows indicate gold particles (LC3) located on the membranes.