Cell Ess allows for protein-protein interaction and transcriptional activation.
A549 cells were stably transfected with a luciferase reporter downstream of NF-kB (NF-kB luc A549) allowing the detection of NF-kB activation. NF-kB A549 cells were grown in media with either FBS or serum free with Cell Ess. TNF-a an activator of NF-kB was added to each culture condition. Luciferase was measured as an indicator of NF-kb activation. Protein- protein interactions (TNF-a and its receptor TNF-R) and NF-kB activation are maintained at similar levels in both Cell Ess and FBS. This activation was shown to be specific NF-kB activity by an inhibitor R0106 targeting NF-kB, which knocked down TNF-a mediated luciferase induction.
The x axis indicates the culture condition for each group. NF-kB-Luc A549 cells were cultured in either 10% FBS in blue or 10% Cell Ess in red. Each of the FBS or Cell Ess groups were then untreated, activated with TNF-a or activated with TNF-a in the presence R0105 (a specific NF-kb inhibitor). Luciferase was measured on the Y-axis as an indicator of NF-kB activity.