JCS003390 Supplementary Material

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• Supplemental Figure 1 -

  Fig. S1. Antiserum to LINT-25 detects recombinant, ectopically expressed and endogenous LINT-25. (A) His-tagged, recombinant LINT-25 was analyzed on a Coomassie-stained gel (Coom) and by western blotting using αHis-Tag reagent (αHis) or polyclonal LINT-25 antiserum (αLINT-25). (B) GFP- or Myc-tagged LINT-25 was transiently expressed in HeLa cells and samples were processed for western blotting 24 hours after transfection using antibodies to the specific tags (αGFP, αMyc) or affinity purified LINT-25 antiserum (αLINT-25). Lysates of mock-transfected HeLa cells were used as negative control (mock). (C) Whole-cell lysates from different human cell lines were analyzed by western blotting using purified LINT-25 antiserum. Recombinant LINT-25 was loaded as positive control (rec. LINT-25). SW480: colon cancer cell line, HEK, human embryonic kidney cell line; HeLa, human epithelial cervix carcinoma cell line (D) HeLa cell nuclei (right panel) or HeLa cells (left panel) were lysed in the presence or absence of dithiothreitol (DTT) and iodoacetamide (IAA). Samples were analyzed by immunoblotting using LINT-25 antiserum. Numbers indicate molecular masses of marker proteins in kDa; mLINT-25, monomeric LINT-25; dLINT-25, dimeric LINT-25; La, lamin.