Figure 2. Localization of wild-type and mutant H1GFP.

(A) H1GFP (84 kDa) expression following transfection into HeLa cells was compared with endogenous IMPDH (55 kDa) by Western blot with anti-GFP antibodies and anti-IMPDH serum (26 h post-transfection shown). Additional experiments confirm that detection is linear within this range (results not shown). (B) Live fluorescence localization of wild-type (wt), R224P or V268I H1GFP protein in HeLa cells shown as GFP fluorescence. (C) The ratio of nuclear to cytoplasmic H1GFP. As described in the Experimental section, the mean GFP fluorescence intensities for the nucleus and cytoplasm were determined for >20 cells of each type. The results are means±S.D., and those significantly different from wild-type are denoted by an asterisk (*P<1×10⁻⁵ and 0.001 for R224P and V268I respectively).