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 (lane 1) or transfected with FLAG-p73 DNA (lanes 2 and 3). Overexpression of FLAG-p73 can be specifically blocked by co-transfection with p73 siRNA (lane 3). FLAG-p73 was detected using p73 Antibody #4662, and p42 was detected using p42 MAPK Antibody #9108. p73 Antibody confirms silencing of exogenous p73 expression, and the p42 MAPK Antibody is used to control for loading and siRNA specificity.
CST recommends transfection with 50-100 nM p73 siRNA 48 hours prior to cell lysis. See protocol for transfection procedure.
SignalSilence® siRNA is supplied in RNAse-free water. Aliquot and store at -20ºC.
SignalSilence® p73 siRNA from Cell Signaling Technology allows the researcher to specifically inhibit p73 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.
The p53 family member, p73, exists in multiple isoforms/splice variants and can induce apoptosis and cell cycle arrest in response to DNA damage via its activity as a transcription regulator (1-3). Upon DNA damage, p73 is phosphorylated at Tyr99 by c-Abl, causing translocation to the nuclear matrix (4). DNA damage-induced acetylation of p73 at Lys321 by the acetyltransferase p300 has also been reported to enhance transcription of genes including that of p53AIP1 (5). Another report, however, indicates that p300 does not acetylate full-length p73 in vivo (6).
RNA interference has been used to block p73 expression in SW480 cells, resulting in chemoresistance (7).
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