Fig 2: TRIM25 ubiquitinates RIG-I through direct interaction. (A) RIG-I directly interacts with TRIM25. Flag-RIG-I and HA-TRIM25 plasmids were co-transfected into HEK 293T cells, and the cells were collected 36 h later. The cell lysates were incubated with conjugated magnetic beads loaded with normal IgG or anti-FLAG antibody at 4 °C for 10 h. The collected cell lysates and IP products were subjected to Western blot analysis with anti-Flag antibody and anti-HA antibody. (B) TRIM25 ubiquitinates RIG-I. HEK 293T cells were transfected with Flag-RIG-I, Myc-Ub, and HA-TRIM25 plasmids for 36 h. Western blots of whole-cell lysates or Flag immunoprecipitates were analyzed using specific antibodies.

Fig 3: Effects of DHM on MM tumor growth and EMT in xenograft mice by promoting the STAT1/RIG-I pathway. (A) Detection of the body weights of xenograft mice, as well as MM tumor weights and volumes in xenograft mice. (B) Detection of protein levels of E-cadherin, N-cadherin, p-STAT1/STAT1, and RIG-I in xenograft mice by western blotting. *p < 0.05 and **p < 0.01 vs. Control group. DHM, dihydromyricetin; MM, multiple myeloma; EMT, epithelial-mesenchymal transition.

Fig 4: A model for the suppressive effect of TRIM25 on RABV replication by activating the RIG-I-IFN axis.

Fig 5: Rig-I knockout combined with PD-1 monoclonal antibodies can be used to treat solid tumours that are insensitive to PD-1 antibodies.(A) Rig-I+/+ and Rig-I−/− mice were inoculated subcutaneously with MC38 cells and treated with a control immunoglobulin (cIg) or PD-1 monoclonal antibody, respectively. (B) Growth curve of MC38 tumours. (C) Anatomy of MC38 tumours. (D) Rig-I+/+ and Rig-I−/− mice were inoculated subcutaneously with B16F10 cells and treated with cIg or a PD-1 monoclonal antibody, respectively. (E) Growth curve of B16F10 tumours. (F) Anatomy of B16F10 tumours. (G, H) Survival curves of immunodeficient mice with MC38 (G) and B16F10 (H) tumours in which Rig-I+/+ or Rig-I−/− naïve CD8+ T cells were transferred alone, treated with anti-PD-1 antibodies alone, or treated with their combination. Data information: The data represented different numbers (n = 5 or 6 for B and E, n = 10 or 11 for G and H) of biological replicates, and were shown as the means ± SEMs. Two-way ANOVA was used in (B, E) (Exact p values were reported on graphs). The log-rank test was used in (G, H) (Exact p values were reported on graphs). *P < 0.05, **P < 0.01, ***P < 0.001, and NS not significant compared with the other groups. Source data are available online for this figure.