Fig. 7: PRMT9-mediated MAVS methylation inhibits MAVS aggregation and activation.
From: The protein arginine methyltransferase PRMT9 attenuates MAVS activation through arginine methylation
a SDD-AGE analysis (top) and SDS-PAGE (below) of the aggregation of MAVS in HEK293T cells cotransfected with HA-MAVS, GFP-PRMT9 and PRMT9 (G260E). b SDD-AGE and SDS-PAGE analysis of the aggregation of MAVS in Prmt9CKO and Prmt9WT peritoneal macrophages cells infected with SeV for 0–8 h. c SDD-AGE and SDS-PAGE analysis of the aggregation of MAVS in Prmt9CKO and Prmt9WT cells reconstituted with empty vector (− or Vec) or mPRMT9 (WT) or mPRMT9 (G260E) infected with SeV for 8 h. d qRT-PCR analysis of Ifnb1 mRNA in Prmt9CKO and Prmt9WT reconstituted with empty vector or mPRMT9 (WT) or mPRMT9 (G260E) infected with SeV or VSV for 8 h, mRNA results are presented relative to those of untreated cells transfected with a Vector plasmid (Average of three replicates), left panel: *P = 0.0130, #P = 0.0241, ns = 0.6343 in sequence; right panel: *P = 0.0105, ##P = 0.0070, ns = 0.4865 in sequence. e Co-IP analysis of SDMA of MAVS in MAVS–/– HEK293T cells cotransfected with HA-MAVS (WT) or HA-MAVS (R41K, R43K) in the presence of GFP-PRMT9 expression plasmid. f SDD-AGE and SDS-PAGE analysis of the aggregation of MAVS in MAVS–/– HEK293T cells transfected with HA-MAVS (WT) or HA-MAVS (R41K, R43K) in the presence of GFP-PRMT9 expression plasmid infected with SeV for 8 h. g qRT-PCR analysis of IFNB (*P = 0.0224, ns = 0.4906 in sequence), IFNA4 (***P = 0.0006, ns = 0.5296 in sequence) and CXCL10 (**P = 0.0043, ns = 0.3199 in sequence) mRNA treated as in f, mRNA results are presented as in d. h SDD-AGE and SDS-PAGE analysis of the aggregation of MAVS with the crude mitochondrial extracts prepared from MAVS–/– HEK293T cell after HA-MAVS and HA-MAVS (R41K, R43K) transfection, followed by incubation in the in vitro aggregation buffer with E1, 5’ppp RNA, GST-PRMT9 and ubiquitin. qRT-PCR data in d, g represent the mean ± SD (n = 3 independent experiments). The data shown in a–c and e–h are from one representative experiment of at least 3 biological independent experiments. *P < 0.05, **P < 0.01, #P < 0.05, ##P < 0.01, ns, not significant (Two-tailed Student’s t-test).
