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. 2017 Jul 18;7:5676. doi: 10.1038/s41598-017-06076-4

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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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Inhibition of exophagy of ANXA2 reduces ANXA2-mediated efferocytosis. Jurkat T cells were treated with 50 μM etoposide for 24 h to induce cell apoptosis. ATG5, RAB8A, RAB27A, or RAB27B knockdown or pcDNA-RAB11wt- and pcDNA-RAB11S25N-overexpressing A549 cells were treated with 500 U/ml IFN-γ for 48 h. After treatment, these A549 cells were co-cultured with apoptotic Jurkat T cells for 2 h. Cells were then washed, fixed, permeabilized and stained with DAPI and rhodamine-phalloidin for nucleus and F-actin, respectively. Cells were mounted and analyzed by confocal microscopy. The nuclei of apoptotic T cells were indicated by white arrowheads in ATG5 knockdown cells (a), pcDNA-RAB11wt- and pcDNA-RAB11S25N-overexpressing cells (c), and RAB8A, RAB27A, RAB27B knockdown cells (e). Scale bar: 10 μm. Cells containing nuclei fragments were quantified in ATG5 knockdown cells (b), pcDNA-RAB11wt- and pcDNA-RAB11S25N-overexpressing cells (d), and RAB8A, RAB27A, RAB27B knockdown cells (f). Data are represented as mean ± SD. ***P < 0.001. At least 70 cells were counted in each treatment group.