Expression of EBNA-3 (B, E, H – red) from recombinant vaccinia virus in cells that were transfected with GFP-F538 (A, D, G – green) leads to the accumulation of GFP-F538 in the nucleus in parallel with the redistribution of EBNA-3 from homogeneous nucleoplasmic pattern to well circumscribed nuclear granules. In these granules EBNA-3 (H) shows high levels of co-localization (I) with GFP-F538 (G). Expression of EBNA-5 (C – red) from recombinant vaccinia virus does not change the cytoplasmic localization of GFP-F538 (C – green). Immunofluorescence staining of endogeneous F538 protein with rabbit polyclonal antibodies shows cytoplasmic distribution in the EBV negative BL cells DG75 (J) but gives predominantly nuclear staining in EBV positive BL cells Raji (K) or EBV transformed lymphoblastoid cell line 940110 (L). DG75 cells that express EBNA-5 show the same cytoplasmic distribution as the parental cells (M) whereas EBNA-3 expressing DG75 cells demonstrate nuclear accumulation of the F538 protein (N). High magnification image of the nucleus show that F538 preferentially accumulates in low DNA density areas that corresponds to the euchromatin (O) DNA staining with Hoechst 33258 is blue.
Kashuba et al. BMC Cell Biology 2002 3:23 doi:10.1186/1471-2121-3-23