Figure 3. OGT deficiency induces cGAS/STING-dependent the type I IFN pathway.
(A–D) Real-time PCR analysis of cytokines mRNA expression in different Ogt−/− cell lines including MC38 (A), LLC (B), HT29 (C), B16-OVA (D) cells. (E) Western blot analysis of the activation of the interferon signaling pathway in different Ogt−/− cell lines including MC38, LLC and B16-OVA cells. (F) Western blot analysis of the activation of the interferon signaling pathway in Ogt−/− rescued cell lines including MC38, LLC, HT29 and B16-OVA cells. (G–H) Real-time PCR and western blot analysis of cytokines mRNA expression and the activation of the interferon signaling pathway in different Ogt−/−Mavs−/− double knockout clones in MC38 cells. (I–K) Real-time PCR and ELISA analysis of cytokines mRNA expression in Ogt−/−cGAS−/− double knockout clones in MC38 (I–J), HT29 (K) cells. (L) Western blot analysis of the activation of the interferon signaling pathway in Ogt−/−cGAS−/− or Ogt−/−Sting−/− double knockout clones in MC38, HT29 and B16-OVA cells. (M–N) BMDCs pre-treated with B16-OVA-Ogt−/− (L), B16-OVA-Ogt−/−cGAS−/− or B16-OVA-Ogt−/−Sting−/− cells (M) supernatant, and co-cultured with OT-1 T cell, then T cell proliferation was evaluated by flow cytometry, OVA257-264 as a positive control. Representative fluorescence-activated cell sorting histograms and statistical data are shown. Data are representative of two or three independent experiments. Statistical significance was determined by unpaired Student’s t-test, one-way ANOVA, two-way ANOVA, *p<0.05, **p<0.01, ***p<0.001, ns, no significant difference. Data represent the mean of ± SD.
Figure 3—figure supplement 1. In vitro Cross-Priming of T Cells by Ifnar-/- BMDCs.
