Journal: Oncoimmunology

Article Title: MerTK-mediated efferocytosis promotes immune tolerance and tumor progression in osteosarcoma through enhancing M2 polarization and PD-L1 expression

doi: 10.1080/2162402X.2021.2024941

Figure Lengend Snippet: UNC2025 inhibited M2 polarization of macrophages and enhanced T cell cytotoxic function in mouse osteosarcoma model. FCM analysis of tumor-associated macrophages from the tumor single cell suspensions from mice bearing K7M2 tumors treated with UNC2025 for 14 days. (a) Gating strategy for macrophage identification using myeloid-focused antibody panels. Each data point represented a biological replicate. (b and d) The frequency of tumor-associated macrophages (CD11b + F4/80 + ) in the total live cells. (c, e, and f) The frequency of M2 macrophages (CD11b + F4/80 + CD206 + ) and M1 macrophages (CD11b + F4/80 + CD80 + ) in total CD11b + F4/80 + cells. (g) Gating strategy for lymphocyte identification using lymphocyte-focused antibody panels. Each data point represented a biological replicate. (h and l) The frequency of CD45 + CD3 + T cells in total live cells. (i, m, and n) The frequency of CD8 + T cells and CD4 + T cells in CD45 + CD3 + T cells. (j and o) The frequency of PD-1 + TIM3 + T cells in CD3 + CD8 + T cells. (k and p) The frequency of CD25 + T cells in CD3 + CD4 + T cells. * p < .05; ** p < .01; NS meaning no significant difference (Student’s t -test).

Article Snippet: Anti-mouse monoclonal antibodies directly conjugated with fluorescein were used for flow cytometry: anti-F4/80 FITC, anti-CD11b PC7, anti-CD206 APC, and anti-CD80 PE (BD Pharmingen, USA) and anti-CD16/CD32, anti-CD45 mCHERRY, anti-CD3 FITC, anti-CD4 APCA750, anti-CD8 APC, anti-PD1 PE, anti-TIM3 PC7, and anti-CD25 BV450 (BioLegend, USA).

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