Fig 1: HP1-binding to KDM2A through a nucleolar localization sequence (NoLS-2).(A) An expression vector encoding Flag-HP1a, HP1?, or HP1ß, or the empty vector was cotransfected with an expression vector encoding KDM2A, SF KDM2A (amino acids 543-1162), or GFP fusion protein with the KDM2A fragment (amino acids 667–1162) to 293T cells. Cell lysates were immunoprecipitated with anti-Flag antibody-conjugated agarose and analyzed by Western blotting with anti-KDM2A antibody (ab99242; Abcam) and anti-Flag antibody. One tenth of input samples were also analyzed. (B) An expression vector encoding GFP fusion protein with KDM2A fragments, which were gradual deletions of NoLS-2, was cotransfected with an expression vector encoding Flag-HP1? or the empty vector in 293T cells. Cell lysates were analyzed as described in A. (C) The GFP fusion proteins in (B) were expressed in MCF-7 cells and subcellular localizations of GFP (Green), nucleolin (Red), and nuclei (Blue) were observed. Scale bar corresponds to 10 µm.