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Figure 1 | Virology Journal

Figure 1

From: Characterization of the latent membrane protein 1 signaling complex of Epstein-Barr virus in the membrane of mammalian cells with bimolecular fluorescence complementation

Figure 1

BiFC with LMP1 cytoplasmic domain and TRAFs. HEK-293T cells were transfected with the indicated LMP1 + TRAF pairs and transfection control, pmCherry. NYFP-CTAR1/2 expresses the N-terminus of YFP (NYFP) fused to the cytoplasmic domain of LMP1, amino acids 184-386, at the N-terminus. CYFP-TRAF2 and CYFP-TRAF3 express the CYFP fused to TRAF2 and TRAF3, respectively, at their N-termini. Twenty-four hours post-transfection cells were fixed and examined by fluorescence microscopy for transfection (mCherry expression) in the red panels and fluorescence complementation in the green panels (panels A, B, C, D, E, and F). Representative phase contrast and fluorescence micrographs are shown. High resolution image of NYFP-CTAR1/2 + CYFP-TRAF2 cells were counter stained with DAPI and acquired using a confocal microscope (panel G). In parallel, cells were assayed fourty-eight hours post-transfection by western blotting for protein expression. Lysates were blotted with GFP monoclonal (recognizes the CYFP domain), LMP1, TRAF2, and TRAF3 antibodies (panel H). Lanes 1-6 correspond to the combinations in panels A-F, respectively. Locations of molecular weight markers are indicated. Fifty micrograms of protein were loaded in each lane.

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Virology Journal

ISSN: 1743-422X