Extended Data Fig. 9: iBFAR2 suppresses tumor growth with low cellular and tissue toxicity.

a, Flow cytometric analysis of proliferation and IFNγ-producing CD8⁺ T cells of mouse splenic primary CD8⁺ T cells that were treated with different concentration of iBFAR2 as indicated and stimulated by plate-bound anti-CD3 plus anti-CD28 (α-CD3/28; 1μg/ml) for 3 days. b, Tumor growth of WT mice that were injected s.c. with MB49 tumor cells and then treated with different concentration of iBFAR2 as indicated or vehicle once every 2 days starting from day 7 (n = 9 mice/group). c, Tumor growth of WT mice that were injected s.c. with Py8119 tumor cells (n = 7 mice/group) or B16-F10 melanoma (n = 6 control, n = 8 mice iBFAR2) and then were treated with iBFAR2 (10 mg/ml) or vehicle every 2 days starting from day 7. d, Flow cytometric analysis of tumor-infiltrating CD8⁺ TRM cells, IFNγ-producing CD8⁺ TRM cells, IFNγ-producing and TNFα-producing CD8⁺ T cells as indicated in MB49 tumor-bearing mice that treated with iBFAR2 (10 mg/kg) or vehicle (n = 4 mice/group). e, Immunoblot of phosphorylated (p-) and total STAT1 in MB49 tumor cells. f, The MB49 tumor cells were treated with DMSO or iBFAR2 for the indicated time points based on CCK8 assay. g, Tumor growth of Rag1^−/−mice that were i.v. injected with young and aged mice derived splenic CD8+ T cells and s.c. injected with MB49 tumor cells, and then treated with vehicle (Veh) or iBFAR2 (20 mg/kg) once every 2 days starting from day 9 (n = 5 mice). h, Flow cytometric analysis of apoptosis of mouse splenic primary CD8⁺ T cells that were treated with different concentration of iBFAR2. i, qPCR analysis of the mRNA expression of Il1b and Tnf of different tissues from the tumor-bearing mice. Bar graphs are presented as mean ± s.e.m. A two-tailed Student’s t-test was performed for comparisons. The data are representative of two (b, c, g, i) or three independent experiments (a, d, e, f, h).

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