Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.
Western blot analysis of extracts from HeLa cells, untransfected or transfected with SignalSilence® eIF4B siRNA #6416, using Phospho-eIF4B (Ser422) Antibody (upper) or eIF4B Antibody #3592 (lower). 48 hours following transfection, cells were treated with Rapamycin (50 nM) and U0126 (10 uM) as indicated.
Western blot analysis of extracts from HeLa cells, transfected with either nonspecific control siRNA or eIF4B siRNA. eIF4B was detected using eIF4B Antibody #3592, and eIF4E was detected using eIF4E Antibody #9742. The eIF4B Antibody confirms silencing of eIF4B expression, and the eIF4E Antibody is used to control for loading and siRNA specificity.
CST recommends transfection with 100 nM eIF4B siRNA 48 hours prior to cell lysis. See protocol on datasheet for transfection procedure.
SignalSilence® siRNA is supplied in RNAse-free water. Aliquot and store at -20ºC.
SignalSilence® eIF4B siRNA from Cell Signaling Technology allows the researcher to specifically inhibit eIF4B expression using RNA interference, a method in which gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products are rigorously tested in-house and have been shown to reduce protein expression in specified cell lines.
Eukaryotic initiation factor 4B (eIF4B) is thought to assist the eIF4F complex in translation initiation. In plants, eIF4B is known to interact with the poly-(A) binding protein, increasing its poly-(A) binding activity (1). Heat shock and serum starvation cause dephosphorylation of eIF4B at multiple sites with kinetics similar to those of the corresponding inhibition of translation, while phosphorylation of eIF4B following insulin treatment correlates well with an observed increase in translation (2-5). Multiple kinases, including p70 S6 kinase, can phosphorylate eIF4B in vitro, and at least one serum-inducible eIF4B phosphorylation site is sensitive to rapamycin and LY294002 (6). Recently, Ser406 was identified as a novel phosphorylation site regulated by mitogens (7), and the phosphorylation of this site is dependent on MEK and mTOR activity (7). This phosphorylation is shown to be essential for the translational activity of eIF4B (7).
RNAi has been used to silence eIF4B expression, resulting in cell death (7).
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