Akt induces PHF20 translocation from the nucleus to the cytoplasm.
A, phosphomimic PHF20-S291D but not nonphosphorylatable PHF20-S291A preferentially binds to Akt. MCF7 cells were transfected with indicated plasmids, immunoprecipitated (IP) with FLAG antibody, and detected with Akt antibody (top). Middle and bottom panels show the expression of the transfected plasmids. IB, immunoblot. B, PHF20 localizes in the nucleus, and Akt induces PHF20 nuclear-cytoplasm translocation. MCF7 cells were transfected with FLAG-PHF20 together with/without Red-myr-Akt. After 48 h, cells were stained with FITC-conjugated FLAG antibody and DAPI. C, PI3K or Akt inhibitors but not MEK inhibitor blocked IGF1-induced PHF20 nuclear-cytoplasm translocation. MCF7 cells were transfected with GFP-PHF20. After 36 h, cells were deprived from serum for 12 h and treated indicated drugs for 30 min prior to IGF1 stimulation for 1 h. Cells were stained with Akt antibody and Texas Red-conjugated secondary antibody.