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. 2020 Sep 28;9(9):e1177. doi: 10.1002/cti2.1177

Figure 6.

Figure 6

3M‐052 acts indirectly on tumor cells through cytokine production from DCs. (a) 4T1.2 cells were treated with various IFN agonists for 48 h and assessed for MHC‐I (H2‐Kd) expression. Poly I:C‐transfected cells were compared with Lipofectamine‐treated control cells; (b) BMDCs and (c) BM macrophages were stimulated for 24 h with various IFN inducers and assessed for MHC‐I (H2‐Kd) expression by flow cytometry and production of (d) IFN‐β by enzyme‐linked immunosorbent assay and (e) IL‐6, (f) IL‐1β and (g) TNF‐α by cytometric bead array (CBA). Conditioned medium (s/n) from 3M‐052‐treated (h) 4T1.2, (i) BMDCs and (j) BM macrophages was transferred onto fresh 4T1.2 cells ± IFNAR1 blocking mAb, MAR1‐5A3. Following 48 h, recipient 4T1.2 MHC‐I (H2‐Kd) expression was assessed by flow cytometry. BM MΦ: bone marrow‐derived macrophages. MFI: mean fluorescence intensity. n = 3–6 biological replicates/group. Experiments were independently repeated twice, and representative data are shown. All data n = 3 replicates/group except (d) n = 6 replicates/group. Statistical analysis was performed by one‐way ANOVA with post hoc Tukey's multiple comparison test. Error bars are SEM. *P < 0.05; **P < 0.01; ***P < 0.001; and ****P < 0.0001.