Fig. 2. Intranuclear localization and expression of the EGFP-WRN fragments shown in Fig. 1A. HeLa cells were transfected with the following constructs: (A) EGFP-WRN (1-1432) (a-d); EGFP-WRN (54-946 +NLS) (e-h); EGFP-WRN (949-1432) (i-l); (B) EGFP-WRN (NLS) (a-d); EGFP-WRN (1072-1432) (e-h); and EGFP-WRN (949-1092 +NLS) (i-l). After transfection, the localization of each construct was analyzed by confocal microscopy. Two representative pictures of each construct are shown. (C) EGFP-WRN (949-1092 +NLS; green)-transfected cells were fixed, and probed with anti-B23 antibody (Texas Red). (D) Protein expression of the different EGFP-WRN constructs. Equal amounts of protein from EGFP-WRN-construct-transfected cell lysates were loaded in each lane. The asterisks correspond to the crossreactivity of the anti-GFP antibody. Arrows show the expression of the GFP fusion protein.