Fig. 3: Oncogenic KRAS significantly suppresses type I IFNs via global epigenetic control.
A WiDr cells were infected with lentivirus carrying pBabe-puro-vector (Vec.) or pBabe-puro-KRASG12D and selected for three days. WiDr-Vec. and WiDr-KRASG12D cells were irradiated (5 Gy), and then harvested for qRT-PCR analysis after 24 hr (n = 3). One-way ANOVA t test. *p < 0.05. B CoLo320 cells were infected with lentivirus carrying pBabe-puro-vector (Vec.) or pBabe-puro-KRASG12D and selected for three days. CoLo320-Vec. and CoLo320-KRASG12D cells were irradiated (5 Gy), and then harvested for qRT-PCR analysis after 24 hr (n = 3). One-way ANOVA t test. *p < 0.05. C HCT116 cells were infected with lentivirus carrying pLKO-scramble shRNA (shNC) or pLKO-shKRAS (shKRAS) and selected for three days. HCT116shNC and HCT116shKRAS cells were irradiated (5 Gy), and then harvested for qRT-PCR analysis after 24 hr (n = 3). One-way ANOVA t test. *p < 0.05 and **p < 0.01. D SW620shNC and SW620shKRAS cells were irradiated (5 Gy), and then harvested for qRT-PCR analysis after 24 hr (n = 3). One-way ANOVA t test. *p < 0.05 and **p < 0.01. E WiDr and Colo320DM (endogenous wild-type KRAS) cells were individually infected with vector (Vec.) and KRASG12D. Cells were selected by puromycin for three days. The level of DNMT1, DNMT3a and DNMT3b was evaluated by immunoblotting (n = 3). One-way ANOVA t test. **p < 0.01. F CT26, HCT116 and SW620 (endogenous mutant KRAS) cells individually infected with lentivirus carry shNC and shKRAS. Cells were selected by puromycin for three days. The level of DNMT1, DNMT3a and DNMT3b was evaluated by immunoblotting (n = 3). One-way ANOVA t test. **p < 0.01. G KRASG12D-tranduced WiDr (endogenous wild-type KRAS) cells were treated with 2.5 μmol/L and 5.0 μmol/L AZA for two days. The level of STING was evaluated by immunoblotting (n = 3). One-way ANOVA t test. *p < 0.05 and **p < 0.01. H. KRASG12D-tranduced CoLo320 (endogenous wild-type KRAS) cells were treated with 2.5 μmol/L and 5.0 μmol/L AZA for two days. The level of STING was evaluated by immunoblotting (n = 3). One-way ANOVA t test. *p < 0.05 and **p < 0.01. I HCT116 and SW620 (endogenous mutant KRAS) cells individually infected with lentivirus carry shDNMT1, shDNMT3a and shDNMT3b. Cells were selected by puromycin for three days. The mRNA level of STING was evaluated by qRT-PCR (n = 3). One-way ANOVA t test. *p < 0.05. J The protein level of STING was evaluated by immunoblotting (n = 3).
