Normal Cellular Expression and Localization of Repair-Deficient XPC Mutants

<div><p>(A) Immunoblot analysis of XP–C fibroblasts transfected with pcXPC vectors coding for wild-type protein or repair-deficient mutants. Soluble cell lysates (20 μg) were separated on polyacrylamide gels and probed for XPC protein using a specific monoclonal antibody directed against the C-terminal sequence of human XPC. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was recorded as the internal standard. Lane 2: XP–C fibroblasts transfected with the pcDNA control vector.</p> <p>(B) Densitometric quantification of three to five independent experiments. The intensity of immunoreactive bands corresponding to XPC protein was normalized against the glyceraldehyde-3-phosphate dehydrogenase standard and reported as the percentage of the wild-type signal (± SD).</p> <p>(C) Immunoblot analysis of XP–C fibroblasts transfected with pGFP (lane 1), or the vectors coding for wild-type (lane 2) or mutant XPC proteins (lanes 3–7) fused to GFP. The primary antibody was directed against GFP.</p> <p>(D) Time course of fluorescent fusion protein expression in XP–C fibroblasts transfected with pXPC–GFP containing the wild-type sequence. Nuclei were stained with Hoechst reagent.</p> <p>(E) Distribution of GFP in XP–C fibroblasts.</p> <p>(F and G) Representative images demonstrating the nuclear localization of repair-deficient mutants 15 h after transfection.</p>