Fig 1: C100-induced STING activation requires nuclear DNA damage(A) BMDCs from WT or Tmem173−/− mice were treated with 8 μg/mL HDCPs for 2 h. Histogram overlay of MitoSOX fluorescence in single live cells. Data are representative of two independent experiments.(B) WT bone marrow precursors were cultured to BMDCs as usual (control [Ctrl]) or in low-dose ethidium bromide (EtBr). Ctrl and EtBr BMDCs were treated with 8 μg/mL HDCPs for 24 h or DMXAA for 3 h. Ifnb mRNA levels were calculated by qPCR with respect to Actb and Rps18. Data are expressed as mean ± SD for technical triplicates with respect to Actb and are representative of 3 independent experiments. Statistical analysis was performed by two-tailed unpaired Student’s t tests with the Holm-Sidak method for multiple comparisons. ∗∗p < 0.01 and ∗∗∗p < 0.001.(C) BMDCs were treated with 5 μg/mL HDCPs for 9 h or 100 μM etoposide for 24 h and then monitored for DNA fragmentation with the APO-BrdU (bromodeoxyuridine) TUNEL kit. Data are representative of two independent experiments.(D) Ctrl and EtBr BMDCs were left untreated or treated with 5 μg/mL C100 for 8 h and then monitored for DNA fragmentation. Data are representative of two independent experiments.(E and F) C57BL/6 mice were injected s.c. on the flank with B16F10 cells. Tumor-bearing mice were randomized and injected i.t. with C100 or intraperitoneally with cisplatin on days 0, 4, and 8. (E) Unpaired two-tailed t test or (F) Mantel-Cox tests were used to determine statistical significance between treatments.∗p < 0.05 and ∗∗p < 0.001. (E and F) n = 7 mice per group.