Figure 1.

Polyubiquitination in Ixodes scapularis. A, Protein lysates were obtained from I. scapularis ISE6 cells and aliquots (10–100 µg) resolved in 10% SDS-PAGE. Dashes represent immunoblots treated with Ub^K63 and Ub^K48 antibodies. Antibody specificity was assessed by preincubating the antibodies with either TetraUb^K48 or TetraUb^K63 for 1 h prior to immunoblotting. B, ISE6 cells, (C) tick salivary glands (SG), and midguts (MG) were fixed with paraformaldehyde and stained with DAPI (blue), Evans blue (red), and FITC (green) Ub^Pan, Ub^K63, or Ub^K48 antibodies. The scale represents 10 µm in (B) and 20 µm in (C). Original magnification at 63× (B) and 20× (C). These experiments were repeated twice. Abbreviations: DAPI, 4′, 6-diamidino-2-phenylindole; FITC, fluoresceine-isothiocyanate; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.