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. 2020 Sep 21;8(2):e001224. doi: 10.1136/jitc-2020-001224

Figure 4.

Figure 4

MDA5 and IFNαβR on stromal cells is required for the antitumor effects and T cell tumor infiltration of poly-IC stabilized with poly-lysine and carboxymethylcellulose (PICLC) therapy. (A, B) The role of MDA5 on stromal cells in the antitumor effect and infiltration. (A) Bone marrow chimeric mice were generated as described in ‘Materials and methods’ section and were inoculated subcutaneously on day 0 with 3×105 B16F10 cells and treated with 50 μg PICLC (intravenously) in combination with αPD-L1 monoclonal antibody (mAb) as described in figure 1. (A) Tumor growth was monitored (data are shown as tumor size±SD; n=2–3 mice per group; ***P<0.001, **p<0.01 by two-way analysis of variance) and (B) the percentages of CD8 T cells and CD11b+/Gr1+ cells were assessed on day 19. Bars are shown as mean±SD; n=2–3 mice per group. **P<0.01 by Mann-Whitney unpaired test. (C) IFN-I production by endothelial cells after poly-IC or PICLC treatment. H5V or bEnd.3 (1×105) mouse endothelial cells were stimulated with PICLC or poly-IC and 24 hours later and the production of IFN-I was assessed in the supernatants. (D) VCAM-I expression in endothelial cells after poly-IC or PICLC treatment. 1×105 bEnd.3 cells in triplicates were stimulated with 50 µg PICLC and the VCAM-I expression was assessed by flow cytometry 24 and 48 hours later. Error bars represent the SD of the mean fluorescence intensity (MFI). **P<0.01 and ***p<0.001 by Mann-Whitney unpaired test.