Figure 1.
In vitro transcription/translation was performed in the presence of [35S] cysteine in a TNT rabbit reticulocyte lysate programmed with either pGEM-4Z-HLTF encoding both HLTFMet1 and Met123 variants (lanes 1–4) or Luciferase T7 control plasmid (lanes 5–8). The products were resolved by electrophoresis on a 4–12% SDS-PAGE gradient and detected by autoradiography. The samples were either 1,5 μl of total in vitro translation products (lanes 2, 4, 6, and 8), or 15 μl of in vitro translation products immunoprecipitated with either the ASE2 antiserum directed against HLTFMet123 (lanes 1 and 5) or the ART2 antiserum directed against HLTFMet1 (lanes 3 and 7). The autoradiography was exposed 3 days. The sizes of the molecular weight markers (MW) are given in kDa. Arrowheads indicate the HLTF proteins.