Cell Signaling Technology

Product Pathways - Akt Signaling

SignalSilence® GSK-3α siRNA #6312

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untransfected or transfected with either GSK-3alpha/beta siRNA #6301 or GSK-3alpha siRNA. GSK-3alpha and GSK-3beta were detected using a GSK-3alpha/beta antibody, and eIF4B was detected using eIF4B Antibody #3592. The GSK-3alpha/beta antibody confirms silencing of GSK-3alpha and beta expression, and the eIF4B Antibody is used to control for loading and siRNA specificity.

Fluorescent Detection

Fluorescent Detection

Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.

Directions for Use

CST recommends transfection with 100 nM GSK-3α siRNA 48 hours prior to cell lysis. See protocol for transfection procedure.

Background

Glycogen synthase kinase-3 (GSK-3) was initially identified as an enzyme that regulated glycogen synthesis in response to insulin (1). GSK-3 is a ubiquitously expressed serine/threonine protein kinase that phosphorylates and inactivates glycogen synthase. GSK-3 is a critical downstream element of the PI3 kinase/Akt cell survival pathway, and its activity can be inhibited by Akt-mediated phosphorylation at Ser21 of GSK-3α and Ser9 of GSK-3β (2,3). GSK-3 has been implicated in the regulation of cell fate in Dictyostelium, and is a component of the Wnt signaling pathway required for Drosophila, Xenopus and mammalian development (4). GSK-3 has been shown to regulate cyclin D1 proteolysis and subcellular localization (5).

Small Interfering RNA (siRNA) has been used to specifically silence GSK-3α expression, causing an upregulation of β-catenin protein levels (6).

  1. Welsh, G.I. et al. (1996) Trends Cell. Biol. 6, 274-279.
  2. Srivastava, A.K. and Pandey, S.K. (1998) Mol. Cell. Biochem. 182, 135-141.
  3. Cross, D.A. et al. (1995) Nature 378, 785-789.
  4. Nusse, R. (1997) Cell 89, 321-323.
  5. Diehl, J.A. et al. (1998) Genes Dev. 12, 3499-3511.
  6. Yu, J. Y. et al. (2003) Mol. Ther. 7, 228-236.

Application References

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Companion Products

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.

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