Additional file 5.
NF-κB activation and kinetics. a) Comparison of NF-κB activation in parental, non-transduced cell lines with p65-GFP-expressing cell lines. Reporter and parental cell lines respond similarly to a NF-κB stimulus. TNFα (0.5 and 10 ng/ml) was added to reporter and parental cell lines for the indicated times. Degradation of IκBα and actin (control) was analyzed by Western blot. Blots are representative of at least three independent experiments. b) Kinetics of p65-GFP nuclear translocation corresponds to IκBα degradation. The reporter cell lines were activated with TNFα (10 ng/ml or 0.5 ng/ml) for the stated time, fixed, stained with Hoechst 33342 and analyzed with automated microscopy and the Scan^R analysis software. For software analysis, individually optimized setups were used for each cell line as shown in Figure 2 and Additional file 3. Error bars = SD of experiment performed in triplicates. Results are representative of three independent experiments.
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Bartfeld et al. BMC Cell Biology 2010 11:21 doi:10.1186/1471-2121-11-21