Fig. 3. Comparison of subcellular distribution of endogenous eIF5A(Hpu), exogenous eIF5A(Lys) and exogenous eIF5A(Hpu).
(A) HeLa cells were fixed after 48 h of transfection, incubated with primary antibodies, eIF5A antibody (NIH353, rabbit polyclonal) or anti FLAG antibody (mouse monoclonal from Sigma) followed by the secondary antibodies, Alexa Fluor 488-conjugated secondary anti-rabbit antibodies (green, Invitrogen) or Alexa Fluor 594-conjugated secondary anti-mouse antibodies (red, Invitrogen) for detection of endogenous eIF5A and exogenous FLAG-eIF5A, respectively. (B) HeLa cells, transfected with pCEFL/GFP-eIF5A without or with vectors encoding DHS and DOHH, were fixed and GFP-eIF5A precursor and GFP-eIF5A(Hpu) were directly visualized. (C) Fluorescence of GFP or GFP-eIF5A formed in HeLa cells transfected as in (B) was visualized by a confocal microscope.
