Product Pathways - PI3K / Akt Signaling
SignalSilence® FoxO1 siRNA I (Mouse Specific) #6468
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Western blot analysis of extracts from C2C12 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® FoxO1 siRNA I (Mouse Specific) (+), or SignalSilence® FoxO1 siRNA II (Mouse Specific) #6493 (+) using FoxO1 (C29H4) Rabbit mAb #2880 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The FoxO1 (C29H4) Rabbit mAb confirms silencing of FoxO1 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.
SignalSilence® FoxO1 siRNA I (Mouse Specific) from Cell Signaling Technology (CST) allows the researcher to specifically inhibit FoxO1 expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.
Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.
Directions for Use
CST recommends transfection with 100 nM SignalSilence® FoxO1 siRNA I (Mouse Specific) 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.
Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.
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).
- Anderson, M.J. et al. (1998) Genomics 47, 187-199.
- Galili, N. et al. (1993) Nat. Genet. 5, 230-235.
- Borkhardt, A. et al. (1997) Oncogene 14, 195-202.
- Nakae, J. et al. (1999) J. Biol. Chem. 274, 15982-15985.
- Rena, G. et al. (1999) J. Biol. Chem. 274, 17179-17183.
- Guo, S. et al. (1999) J. Biol. Chem. 274, 17184-17192.
- Seoane, J. et al. (2004) Cell 117, 211-223.
- Arden, K.C. (2004) Mol. Cell 14, 416-418.
- Yang, Y. et al. (2005) EMBO J. 24, 1021-1032.
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- 6568 SignalSilence® Control siRNA (Unconjugated)
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
- 6493 SignalSilence® FoxO1 siRNA II (Mouse Specific)
- 9454 FoxO1 (L27) Antibody
Limited Use Label License, RNA interference: This product is licensed under European Patent 1144623 and foreign equivalents from Ribopharma AG, Kulmbach, Germany and is provided only for use in non-commercial research specifically excluding use (a) in drug discovery or drug development, including target identification or target validation, by or on behalf of a commercial entity, (b) for contract research or commercial screening services, (c) for the production or manufacture of siRNA-related products for sale, or (d) for the generation of commercial databases for sale to Third Parties. Information about licenses for these and other commercial uses is available from Ribopharma AG, Fritz-Hornschuch-Str. 9, D-95326 Kulmbach, Germany.
For Research Use Only. Not For Use In Diagnostic Procedures.