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. 2023 Jan 10;11(1):e005871. doi: 10.1136/jitc-2022-005871

Figure 6.

Figure 6

CI-994 sensitizes MYC-driven medulloblastoma cells to anti-CD47 phagocytosis checkpoint immunotherapy: (A) Tumor inflammation on CI-994 treatment was evaluated by an ELISA-based assay for IFN-γ secretion. A significant increase (**p<0.05: non-parametric t-test) is seen in MB002, D425, and D283 cell lines on treatment at 2.5 μM CI-994. (B) An increase in expression of surface calreticulin is seen in MB002 (left panel) and D425 (right panel) as assayed by flow cytometry. (C) An increase in the relative surface calreticulin expression per cell was assayed by measuring the MFI for four different cell lines. (D) Levels of HMGB1 were assayed by a standard colorimetric ELISA. (E) D425 cells were incubated with human peripheral blood mononuclear cells-derived macrophages and tumor cells pretreated with either IgG control, CI-994, anti-CD47 mAb (B6H12), or a combination of CI-994+ anti-CD47 mAb. Phagocytosis was assayed using flow cytometry. (F) Survival analysis of D425 tumor-bearing mice treated with either control, CI-994, anti-CD47, or a combination of anti-CD47 and CI-994.(p=0.0002 using the log-rank (Mantel-Cox) test with a HR of 3.669). DMSO, dimethyl sulfoxide; IFN, interferon; mAb, monoclonal antibody; MFI, mean fluorescence intensity.