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. 2018 Jul 19;9:1650. doi: 10.3389/fimmu.2018.01650

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Copyright © 2018 Vidyarthi, Khan, Agnihotri, Negi, Das, Aqdas, Chatterjee, Colegio, Tewari and Agrewala.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Polarization of macrophages to M1, M2a, and M2c phenotypes. Bone marrow cells (BMs) were cultured in the presence of conditioned medium containing M-CSF for 6 days. Later, macrophages were differentiated into M1, M2a, and M2c phenotypes using LPS + IFN-γ, IL-4 + IL-13, and TGF-β + IL-10, respectively. The M1, M2a, and M2c macrophages (F4/80⁺ gated cells) were characterized by the (A) display of CD86, CD40, and PDL-1 through flow cytometry. Number in the inset indicates the MFI; (B) secretion of IL-6, IL-12, and TNF-α estimated in the culture SNs by ELISA; (C) expression of Irg-47, iNOs, Tim-3, and Arg-1 by quantitative RT-PCR. Data shown as mean ± SD are representative of two to three independent experiments (*p < 0.05; **p < 0.01; ***p < 0.001).