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. 2021 May 17;131(10):e146832. doi: 10.1172/JCI146832

Figure 5. ATT-I enhances antigen presentation on tumor cells.

Figure 5

(A) MFI values of H-2Kb and H-2Kb-SIINFEKL (OVA) on the MC38-OVA cells (shNT control, shPsmd4, and shPsmd7) with ATT-I (0, 5, 10, and 30 μM), which were determined by flow cytometry analysis. Data are presented as mean ± SEM and are representative of 2 independent experiments (n = 2). Statistical analysis was conducted using 1-way ANOVA. (B and C) HLA-A,B,C on the surface of HCT116 cells (B) and SW837 cells (C) was analyzed by 3D confocal imaging of immunofluorescence. Cell nucleus was stained by 4′,6-diamidino-2-phenylindole (DAPI). Quantitative data are presented as mean ± SD of 3 to 4 parallel experiments (n = 3–4). Unpaired 2-tailed t test was used for statistical analysis. Scale bars in B and C: 30 μm. (D) The effect of ATT-I on the cytotoxicity of MC38 OVA+ after blocking the MHC-I/TCR interaction using the MHC-I antibody. Specifically, we treated MC38 OVA+ cells with and without 30 μM ATT-I for 48 hours. Rat IgG2a isotype control and anti-mouse MHC class I (H2) were used for blocking the cells overnight. The antigen-specific cytotoxicity was analyzed by flow cytometry. Data are presented as mean ± SEM and are representative of 2 independent experiments (n = 4). Statistical analysis was conducted using 1-way ANOVA. *P < 0.05; **P < 0.01; ***P < 0.001.