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. 2006 Nov 29;81(4):1762–1772. doi: 10.1128/JVI.01859-06

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Copyright © 2007, American Society for Microbiology

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FIG. 6. — Association between JSRV and enJS56A1 Gag proteins. (A) 293T cells were cotransfected with enJS56A1HA-MA and JSRVFLAG-MA. At 48 h posttransfection, cell lysates were immunoprecipitated (IP) and analyzed by SDS-PAGE/Western blotting (WB) as indicated beside each panel. Gag expression was assessed by Western blotting using a JSRV MA antiserum. enJS56A1-JSRV Gag association is evident in lysates from cells cotransfected with both viruses (lane 4). Note that the JSRV MA antiserum is a polyclonal serum and is much more sensitive in Western blotting than the anti-HA or anti-FLAG monoclonal antibody. (B) Supernatants of cells transfected with the indicated plasmids were resolved by SDS-PAGE and immunoblotted with an antiserum towards JSRV CA. Note that JSRVΔpro releases virus particles into the supernatant with an immature Gag (lane 5) because of the lack of a functional protease in this virus. Coexpression of JSRVΔpro and enJS56A1ΔNC2 (lane 8) leads to the release of viral particles with both immature and mature Gag. (C) Supernatants and lysates of cells transfected with the indicated plasmids were resolved by SDS-PAGE and immunoblotted with an antiserum towards JSRV MA. Note that enJS56A1ΔMHR is highly expressed (lane 4) but does not interfere with JSRV exit (lane 6).