Survivin’s Role in Cell Survival
NFkB Pathway Analysis
Assessing the Roles of 178 Kinases in the Cell Cycle
 

RNAi Experimental Steps

1. Order Silencer Validated siRNA to TNF-alpha Receptor (siRNA ID: 42864)
2. Transfect HeLa cells with TNFR siRNA or IL-1beta siRNA (used as negative control)
3. Incubate 48 hours
4. Replate
5. Treat with IL-1alpha or TNFalpha
6. Incubate for 24 hours
7.Detect NFkB by immunofluorescence and stain nuclei with Hoechst
8. Analyze using Cellomics ArrayScan® HCS Reader

 
 
Conclusion: Silencing of TNF alpha receptor eliminates NFkB translocation in response to TNF-alpha, but not IL-1alpha. NFkB translocation is more efficiently stimulated by IL-1alpha when the TNF-alpha receptor is silenced by RNAi.