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. 2021 Oct 26;12:743354. doi: 10.3389/fimmu.2021.743354

Figure 3.

Figure 3

S100A9 treatment promotes the polarization of bone marrow-derived macrophages to M2 in vitro. (A) Mouse (6week, n=4/group) bone marrow single cell suspension was collected through a 70μm filter, and removed red blood cells by red cell lysis solution. BMDMs (M0) were induced with mM-CSF (10 ng/ml, 6 days) in RPMI1640 medium. Further polarization was achieved by adding 50 ng/ml IFN-γ (M1), 10 ng/ml IL-4 (M2), DMSO (0.3%), S100A9 (1 µg/ml) and S100A9 (1 µg/ml)+tasquinimod (10 µmol/ml) for 48 more hours. (B, C) Immunotyping was performed by flow cytometric analysis to detect live cells, macrophage surface markers (F4/80+), M1-MΦ marker (CD86+), and M2-MΦ marker (CD206+). (D) The ratio of CD206+ and CD86+ in different groups. (E, F) Relative mRNA expression of IL10 and TGFβ in different groups. Data represent the mean ± standard error(n=3–5). *P < 0.05, ***P < 0.001. comparisons between groups using one-way ANOVA followed by Dunnett’s multiple comparisons test.