vFLIP binds to and is stabilized by peroxisome-localized MAVS.

<p>(A-B) Co-IP assays using extracts from BCBL-1 cells (A) and 293T cells transfected with vectors expressing Flag-MAVS or deletion mutants together with vFLIP (B). IgG-Hc indicates heavy chain of immunoglobulin. (C) <i>In vitro</i> GST pull-down assay using the indicated purified recombinant proteins. Since GST-vFLIP was poorly detected by anti-GST antibody, the same membrane was re-probed with anti-vFLIP antibody. Asterisks indicate intact GST-fusion proteins. (D) Diagram of MAVS, MAVS-Mito, and MAVS-Pex constructs. (E-F) Co-IP assays using extracts from <i>MAVS</i> KO 293T cells transfected with the indicated MAVS and V5-vFLIP vectors (E) and of <i>MAVS</i> KO BCBL-1 (1A4) reconstituted with GFP, MAVS^Rg1, and MAVS^Rg1-Pex (F). Asterisk indicates heavy chain of immunoglobulin. (G) Immunostaining of <i>MAVS</i> KO 293T cells transfected with the indicated MAVS and V5-vFLIP vectors. Co-localization (yellow spots) of MAVS-Pex and V5-vFLIP is shown in the enlarged inset. (H) Immunoblots of extracts from <i>MAVS</i> KO 293T cells transfected with V5-vFLIP and the indicated MAVS or GFP vectors. Relative vFLIP band intensities normalized by LDH are marked below the blot. (I) Immunoblots of extracts from <i>MAVS</i> KO 293T cells transfected with V5-vFLIP either with or without MAVS-Pex and incubated in complete media and EBSS for an additional 6 h.