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. 2019 Dec 13;9:19061. doi: 10.1038/s41598-019-55503-1

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© The Author(s) 2019

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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*0401 is localized to lysosomes and *0402 to early endosomes during recycling. (A) 3 D confocal microscopy image showing BMDCs from *0401 and *0402 mice stained for DR (MHCII), early endosomal antigen (EEA1) and lysosomal associated membrane protein 1 (LAMP1). (B) Quantification of MHCII integrated intensity from images in (A) using FIJI (NIH). (C) Images were analyzed for MHCII co-localization with (top) EEA1 and (bottom) LAMP1 using Pearson’s co-localization coefficient in ZEN (Carl Zeiss). Zoomed images are demarcated by white boxes. For each condition, >20 individual cells were imaged. Scale bars, 10 µm. Data represent mean ± SEM (n = 3 mice per group). MHCII recycling in BMDCs treated with (D) lysosome inhibitor, Chloroquine, and (E) protease inhibitor, Bortezomib. Data represent mean ± SEM (n = 3 mice per group) from two independent experiments, P < 0.05.