Fig. 5: PRMT9 targets MAVS.

a Luciferase (Lucif) activity assays of IFN-β reporter in HEK293T cells transfected with expression plasmids for cGAS-STING, RIG-IN, MDA5, MAVS, TBK1, or IRF3-5D along with Flag-PRMT9 or control vector (Ctrl) for 24 h (mean ± SD, two-tailed student’s t-test Flag-PRMT9 vs. Ctrl Vector, ns = 0.0959, ****p < 0.0001, ****p < 0.0001, ****p < 0.0001, ns = 0.5018, ns = 0.5552 in sequence; n = 3 independent experiments). b qRT-PCR analysis of IFNB1 mRNA in HEK293T cells transfected with the indicated adaptors with Flag-PRMT9 or control vector (mean ± SD, two-tailed student’s t-test Flag-PRMT9 vs. Ctrl Vector, *p = 0.0446, ***p = 0.0008, **p = 0.0018, ns = 0.8231, ns = 0.1474 in sequence; n = 3 independent experiments). a, b Results are presented relative to those of untreated cells transfected with a Vector plasmid. c Co-IP analysis of the interaction between PRMT9 and adaptors in HEK293T cells cotransfected with GFP-PRMT9 and HA-cGAS, HA-RIG-I, HA-MAVS, HA-TBKI, Myc-MDA5 or Myc-IRF3 in HEK293T cells plasimds. d In vitro analysis of the interaction between PRMT9 and MAVS, using recombinant protein GST-PRMT9 and His-MAVS incubated in vitro. e Confocal analysis of the colocalization of Myc-MAVS (Red) and GFP-PRMT9 (Green) in HEK293T cells cotransfected for 24 h. Scale bars: 5 μm. Intensity profiles of each line was quantified by ImageJ software and drawn by GraphPad Prism 7.0. Myc-MAVS - GFP-PRMT9 colocalization was also quantified using Pearson’s correlation coefficient method and scatter map. f Co-IP analysis of the interaction between PRMT9 and MAVS in mouse peritoneal macrophages infected with SeV for 0–12 h. Densitometric analysis of protein expression levels, bands were normalized with individual actin, line graphs are presented relative to the first lane. *P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant (Two-tailed Student’s t-test). Similar results were obtained from three independent experiments.