Extended Data Fig. 9: RIG-I inactivation by RIP-III ameliorates mtRNA-induced autoimmune responses in mice.
From: Targeting a key disulfide linkage to regulate RIG-I condensation and cytosolic RNA-sensing
a, In situ PLA of RIG-I and MAVS in MEF cells stimulated of mtRNA (2.5 μg/mL, delivered by transfection) for 10 h, followed by treatment with RIP-III (50 µM) for 2 h (left); n = 3. The PLA-detected proximity (PROX) complexes (red dots) were quantified (right). b, Immunoblot (IB) of p-IRF3, p-TBK1 and IRF3 dimerization (native gel) and total IRF3, TBK1 and qPCR analysis of Ifnb1, Cxcl10 and Ccl5 mRNA expression in Rig-IWT/WT and Rig-IC865S/C865S MEFs transfected without (−) or with (+) mtRNA (2.5 μg/mL) for 10 h, followed by treatment with control BSA (−) or RIP-III (50 µM) for 2 h; n = 3; ***P = 0.0006. c,d, Related to Fig. 8o, qPCR analysis was performed to measure the mRNA levels of Il-1b (left), Il-6 (middle) and Tnf (right) in the spleen (c) and kidney (d) from the indicated groups; n = 6; ***P = 0.0006; (d) left: P = 0.0042, right: P = 0.0021. e,f, H&E staining of mice spleen tissues (e) and kidney tissues (f) from Fig. 8o. Data are representative of at least three independent experiments and shown as mean ± s.d. Scale bar, 10 μm (a), 100 μm (e,f). Statistical analysis was performed using a two-tailed Student’s t-test (a–d); **P < 0.01, ***P < 0.001, ****P < 0.0001. Exact P values, source numerical data and unprocessed blots are provided.
