Peroxisomes support human herpesvirus 8 latency by stabilizing the viral oncogenic protein vFLIP via the MAVS-TRAF complex
(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) In vitro 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 MAVS KO 293T cells transfected with the indicated MAVS and V5-vFLIP vectors (E) and of MAVS KO BCBL-1 (1A4) reconstituted with GFP, MAVSRg1, and MAVSRg1-Pex (F). Asterisk indicates heavy chain of immunoglobulin. (G) Immunostaining of MAVS 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 MAVS 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 MAVS 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.