Fig. 6 | Signal Transduction and Targeted Therapy

Fig. 6

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

Fig. 6

The interaction between Mfn2 and MAVS triggers the ubiquitination of MAVS. a, b Co-immunoprecipitation assays of MAVS and Mfn2 in the lung tissues of mice at 4 d post infection and in A549 cells at 12 h post infection. Whole lysates were subjected to immunoprecipitation with MAVS or Mfn2 antibody attached to sepharose. Whole-cell lysates (WCL) and immunoprecipitates were analyzed by immunoblotting with an anti-Mfn2 or MAVS antibody. c–e The levels of MAVS ubiquitination in Mfn2-overexpressed or Mfn2-knockdown A549 cells were measured by western blotting at 12 h post infection. f Ubiquitination levels were measured in HEK293T cells co-transfected with pCDNA3.1-MAVS-HA, pCDNA3.1-Mfn2-Flag, pCMV-UB-Myc, pCMV-Ub(K48R)-myc, or pCMV-Ub(K63R)-myc and infected with H1N1. Whole lysates from HEK293T cells were subjected to immunoprecipitation with anti-Myc antibodies attached to sepharose. WCL and immunoprecipitates were analyzed by immunoblotting with anti-HA, anti-Myc, and anti-Flag antibodies. g, h The interaction between full length and truncations of Mfn2 with MAVS, between full length and truncations of MAVS with Mfn2 was analyzed by co-immunoprecipitation assay in virus-infected HEK293T cells. i The co-localization of full length and truncations of MAVS and Mfn2 in virus-infected HEK293T cells was observed under confocal microscopy. CORT, corticosterone

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