Polarized M1 and M2 macrophages were detached from tissue culture flask using Accumax™ (Nacalai Tesque, Japan). 2 × 105/ml M1 and M2 macrophages were washed with PBS and stained using antibodies for CD11b-PE, CD14-FITC, CD86-FITC, *CD80-APC/H7, CD206-APC and CD200R-PE (Miltenyi Biotec, Germany) (*BD Biosciences, United States) for 10 min at 4 °C in the dark. Antibodies with similar conjugates were stained in separate tubes. The cells were washed with MACS buffer to remove excess antibodies. Fluorescence was measured via flow cytometry using a FACSCanto™ II flow cytometer and FACSDiva software (BD Biosciences, United States). Histograms were plotted using FlowJo™ version 10 software (BD Biosciences, United States).
Copyright and License information: The Author(s) ©2021 Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this
article to respond.