Monoclonal Antibodies Specific for Disease-Associated Point-Mutants: Lamin A/C R453W and R482W
Figure 7
Wild-type and mutant R453W lamin A/C are present in distinct, non co-localizing foci in EDMD nuclei.
(A) Immunofluorescence of two EDMD nuclei. Cells were co-stained with anti Lamin A/C N-18 and anti R453W. Fluorescence intensity of the lamin A/C staining by the two antibodies along the dashed lines in the magnified images of the nucleoplasm (4th image in each row) and of the nuclear rim (last image in each row) are shown above or below the respective images. Areas of magnification (3.5-fold) are indicated by white squares in the merged images of each nucleus. Bar scale, 10 µm. (B) Cellular fractionation of wild-type and EDMD cells. Whole cell lysate (WC), cytoplasmic (C), whole nuclear (N), low-salt soluble nuclear (LS), low-salt insoluble nuclear (LP), high-salt soluble nuclear (HS), and high-salt insoluble nuclear (HP) fractions were separated by 7.5% SDS-PAGE and analyzed by Western blotting with the indicated antibodies.