Figure 3.

Gelsevirine (GS) promotes K48-linked ubiquitination of STING. HEK293T cells were treated with or without GS (10 μM) for extra 2hrs after transfecting with plasmids expressing STING-HA and UB-flag for 24 hrs. Then, immunoprecipitation (IP) and immunoblot (IB) analysis were carried out (A). Raw264.7 was pretreated with or without GS (10 μM) for 6 hrs and then treated with 2’3’-cGAMP (5 μg/ml) for 3 hrs. Then, IP, and IB analyses were performed (B). HEK293T cells were transfected with plasmids expressing STING-HA for 24 hrs. Next, 10 μM GS was added for 2hrs. Then, IP and IB analyses of SUMRF1, SUMRF2, TRIM21, RNF125, and RNF5 were performed (C). Raw264.7 was pretreated with or without GS (10 μM) for 6 hrs and then activated with 2’3’-cGAMP (5 μg/ml) for 3 hrs. The protein expression of TRIM21 was determined by Western blot (D).