Figure 1
Nucleotide sequence and protein expression of fs-α1S. A) Nucleotide sequence of wt-α1S and fs-α1S in the region of the frame-shift. B) The three reading frames of fs-α1S in the region of the frame-shift are shown. Translation of the C-terminal half of α1S is explained by a restart of translation at the indicated ATG codon which is located 25 bases downstream from the termination codon indicated by the asterisk. C) Confocal images (calibatrion bar is 10 microns) show details of the intracellular distribution of the expressed proteins. Cells were transfected with the CD8 cDNA plus wt-α1S or fs-α1S. Cells were incubated with CD8 antibody beads, fixed, and stained with T7 primary/fluorescein-conjugated secondary antibodies. Pixel intensity was converted to a 16-level inverted gray scale with high-intensity pixels in black color. Asterisks show on-focus CD8 antibody beads (diameter 4.5 microns) bound to cells. NT indicates a non-transfected myotube in the same focal plane of the transfected cell. D) Immunoblots using anti T7 antibody of cultures of dysgenic myotubes expressing wt-α1S and fs-α1S. Indicated are 3 of 7 molecular weight markers run in the same gel.