Fig. 7 | Signal Transduction and Targeted Therapy

Fig. 7

From: Novel insights into stress-induced susceptibility to influenza: corticosterone impacts interferon-β responses by Mfn2-mediated ubiquitin degradation of MAVS

Fig. 7

Identification of SYVN1 as the E3 ligase in Mfn2-mediated MAVS ubiquitination. a Co-immunoprecipitation assay of MAVS and SYVN1 in A549 cells. At 12 h post infection, WCL were subjected to immunoprecipitation with MAVS or SYVN1 antibody attached to sepharose. WCL and the immunoprecipitates were analyzed by immunoblotting with an anti-SYVN1 or MAVS antibody. b Co-immunoprecipitation assay of Mfn2 and SYVN1 in A549 cells. WCL were subjected to immunoprecipitation with Mfn2 or SYVN1 antibody attached to sepharose. WCL and immunoprecipitates were analyzed by immunoblotting with an anti-SYVN1 or Mfn2 antibody. c The interaction of MAVS and SYVN1 in Mfn2-knockdown A549 cells were analyzed by co-immunoprecipitation assay at 12h post infection. d, e NP/MAVS/IFN-β protein expressions in SYVN1-knockdown or overexpressed A549 cells were measured by western blotting at 12 h post infection. f–h MAVS ubiquitination levels in SYVN1-knockdown and overexpressed A549 cells were analyzed by western blotting at 12 h post infection. i The truncated variants of Myc-tagged SYVN1 are depicted. j, k The interaction of truncated Myc-tagged SYVN1 variants with Flag-tagged Mfn2 or HA-tagged MAVS in virus-infected HEK293T cells were analyzed by co-immunoprecipitation assays. l The truncated variants of Flag-tagged Mfn2 or HA-tagged MAVS are depicted. m, n The interaction of Myc-tagged SYVN1 with Flag-tagged Mfn2 depletion mutants or HA-tagged MAVS depletion mutants in virus-infected HEK293T cells were analyzed by co-immunoprecipitation assays. CORT, corticosterone

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