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The Ank9 N-terminus is necessary for localization to Golgi- and ER-positive cellular fractions. Lysates of HeLa cells transfected to express GFP-tagged Ank9, Ank9ΔF-box, or Ank947–422 for 4 h or 16 h were subjected to density gradient fractionation. Western blots of nine successive fractions were screened with GFP, GM130, and calreticulin antibodies to confirm the Ank9 proteins’ subcellular trafficking patterns. Results are representative of two experiments with similar results.
