Fig. 3.

Serine 215 phosphorylation of p53 by Aurora-A controls subcellular localization of p53. a H1299 cells transfected with GFP tagged wild-type (WT) or phosphor-mimetic mutant (SD) of p53 for 24 h were fractionated into cytosolic (Cyto) and nuclear (Nuc) fractions and analyzed by immunoblotting with anti-GFP antibody. The purity of cytosolic and nuclear fractions was confirmed by immunoblotting for anti-Hsp90 and anti-PARP antibodies, respectively. WCE whole cell lysates. b H1299 cells transfected with GFP-p53 WT or SD mutant were counterstained for DNA with DAPI (blue), and subcellular localization of GFP-p53 was analyzed by immunofluorescent microscopy. c WT or phosphor-deficient mutant (SA) of GFP-p53 was co-transfected with Flag-tagged WT or kinase dead mutant (KD) of Aurora-A into H1299 cells. Twenty-four hours after transfection, cells were immunostained with anti-flag antibody (red) and counterstained for DNA with DAPI (blue). Subcellular distribution of GFP-p53 was analyzed by by immunofluorescent microscopy