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. Author manuscript; available in PMC: 2021 Aug 14.
Published in final edited form as: Cell Rep. 2021 Jun 22;35(12):109272. doi: 10.1016/j.celrep.2021.109272

Figure 4. Riok3 interacts with RIG-I/MDA5 and promotes their degradation.

Figure 4.

(A and B) Riok3 interacts with both RIG-I and MDA5. The primary macrophages were infected with VSV at MOI = 1 for indicated times, followed by immunoprecipitation using indicated antibodies.

(C) Riok3 inhibits the expression of RIG-I and MDA5 upon VSV infection. Immunoblot analysis of RIG-I and MDA5 in lysates of Riok3+/+ or Riok3−/− macrophages infected with VSV for indicated times.

(D) Riok3 promotes the degradation of RIG-I and MDA5 upon CHX treatment. Immunoblot analysis of RIG-I and MDA5 in lysates of Riok3+/+ or Riok3−/− macrophages treated with CHX (100 mg/mL) for indicated hours after infection with VSV for 2 h.

(E) Riok3 increases the polyubiquitination of RIG-I and MDA5 in 293T cells. Plasmids encoding Myc-RIG-I (or Myc-MDA5) and HA-ubiquitin were co-transfected into HEK293T cells with or without a plasmid encoding Riok3. The RIG-I and MDA5 ubiquitination was monitored by immunoprecipitation.

(F) Riok3 increases the polyubiquitination of RIG-I and MDA5 in PMs. Riok3+/+ and Riok3−/− PMs were infected with VSV for 4 h, followed by immunoprecipitation for detecting the RIG-I and MDA5 ubiquitination.