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. 2015 Apr 10;4:10.3402/jev.v4.26334. doi: 10.3402/jev.v4.26334

Fig. 2.

Fig. 2

LMP1–TRAF2 association at endosomal membranes controls signalling and sorting. (a) Immunofluorescent labelling of endogenous TRAF2 and TRAF3 (all in green) in wtLMP1 (red) transfected HEK293 cells. Lower panel: Immunofluorescent labelling on EBV-negative BJAB cells carrying a tetracycline-inducible (TET-Off) LMP1 expression construct, induced for 24 hours (LMP1 in red) and labelled for endogenous TRAF2 (green). N indicates nucleus. (b) Immunofluorescent labelling of endogenous TRAF2 (green) in HEK293 cells transfected with LMP1 constructs with point mutations in (LMP1 DM) the suspected TRAF-binding sites (red). N indicates nucleus. (c) Reporter assay for LMP1-wt or LMP1-DM NFκB activity. Cell lysates of HEK293 cells transfected for 24 hours with wtLMP1, LMP1-DM, or empty vector (control), together with an NFκB–reporter construct. Error bars represent s.d.; shown is one representative experiment; n=3. (d) Western blotting analysis on TRAF2 and LMP1 protein levels in EBV-infected LCL cell and exosome lysates (left) and in wtLMP1-transfected HEK293 cell and exosome lysates (right). (e) Western blotting analysis on LMP1 protein levels in wtLMP1– or LMP1-DM-transfected HEK293 cell and exosome lysates.