Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

Phospho-FoxO3a (Ser318/321) Antibody #9465

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R Mk (C) Endogenous 97 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  C=Chicken
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-Fox03a (Ser318/321) Antibody detects endogenous levels of Fox03a only when phosphorylated at serine 318/321. The antibody is expected to cross-react with Fox01 when phosphorylated at serine 322/325 based on the peptide sequence.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human Fox03a. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from serum starved and serum-treated NIH/3T3 cells as well as untreated and LY294002/Wortmannin-treated MDA-MB-468 cells, using Phospho-Fox03a (Ser318/321) Antibody (upper) or Akt Antibody #9272 (lower).

Background

The Forkhead family of transcription factors is involved in tumorigenesis of rhabdomyosarcoma and acute leukemias (1-3). Within the family, three members (FoxO1, FoxO4, and FoxO3a) have sequence similarity to the nematode orthologue DAF-16, which mediates signaling via a pathway involving IGFR1, PI3K, and Akt (4-6). Active forkhead members act as tumor suppressors by promoting cell cycle arrest and apoptosis. Increased expression of any FoxO member results in the activation of the cell cycle inhibitor p27 Kip1. Forkhead transcription factors also play a part in TGF-β-mediated upregulation of p21 Cip1, a process negatively regulated through PI3K (7). Increased proliferation results when forkhead transcription factors are inactivated through phosphorylation by Akt at Thr24, Ser256, and Ser319, which results in nuclear export and inhibition of transcription factor activity (8). Forkhead transcription factors can also be inhibited by the deacetylase sirtuin (SirT1) (9).

Phosphorylation sites Ser322 and Ser325 of Fox01 are important for nuclear export and are homologous to Ser318 and Ser321 of Fox03a (9).

  1. Anderson, M.J. et al. (1998) Genomics 47, 187-199.
  2. Galili, N. et al. (1993) Nat. Genet. 5, 230-235.
  3. Borkhardt, A. et al. (1997) Oncogene 14, 195-202.
  4. Nakae, J. et al. (1999) J. Biol. Chem. 274, 15982-15985.
  5. Rena, G. et al. (1999) J. Biol. Chem. 274, 17179-17183.
  6. Guo, S. et al. (1999) J. Biol. Chem. 274, 17184-17192.
  7. Seoane, J. et al. (2004) Cell 117, 211-223.
  8. Arden, K.C. (2004) Mol. Cell 14, 416-418.
  9. Yang, Y. et al. (2005) EMBO J. 24, 1021-1032.
  10. Rena, G. et al. (2002) EMBO J. 21, 2263-2271.

Application References

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