Extended Data Fig. 3: Teniposide and paclitaxel induce micronuclei formation and anti-tumor immunity to B16F10 tumors in a type I IFN signaling-dependent manner.
From: The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING–IFN-β pathway
a, b, (Left) Immunoblot and (right) in vitro cell growth of (a) MCA205 and (b) B16F10 transduced with indicated shRNA. c, Flow cytometry of tumor-infiltrating CD8+T cells and CD103+CD8α+DCs in MCA205 tumor suspensions isolated from WT or Ifnar1-KO mice on Day 6 (n = 6 per group). d, Confocal fluorescence microscopy of MCA205 stably expressing cGAS-Clover. Scale bar, 10 μm. e, qRT-PCR of Ifnb1 in unstimulated MCA205 and B16F10 transduced with indicated shRNA. f, (Left) Immunoblot and (right) qRT-PCR of Ifnb1 in shDapk3#1-transduced MCA205 ectopically expressing V5-tagged DAPK3(WT) or DAPK3(D161A). Cells were stimulated with 2′,3′-cGAMP, 3′,3′-cGAMP, c-di-GMP (20 μg/ml for all three agonists) or DMXAA (50 μg/ml) for 4 h. g, Confocal fluorescence microscopy of B16F10 stably expressing cGAS-Clover. Cells were treated with teniposide (10 μM) for 24 h or paclitaxel (100 nM) for 72 h. Scale bar, 10 μm. h, (Left) Apoptosis measured in shRNA-transduced B16F10 treated with teniposide (10 μM) for 24 h or (right) paclitaxel (100 nM) for 48 h. i, Tumor volume of B16F10 subcutaneously transplanted into WT and Ifnar1-KO mice and treated with teniposide or paclitaxel (n = 6 for vehicle, n = 7 for teniposide and paclitaxel). Tumor size on Day 15 is represented (right panel). Data are representative (a-d, g-i) or the mean (a, b, e, f) of three independent experiments. Values represented mean ± s.d. *P < 0.05, **P < 0.01, and ***P < 0.001. Statistical comparisons were conducted using two-tailed t-test (a-c, e, f, h, i).
