HT-29 and Caco-2 Reporter Cell Lines for Functional Studies of Nuclear Factor Kappa B Activation

Figure 8

Natural cyclic peptide effect on NF-κB activation induced by TNF-α and LPS on Caco-2-NF-κB-hrGFP C3 and HT-29-NF-κB-hrGFP E5 and F6 clones cells. The natural cyclic peptide was added to Caco-2-NF-κB-hrGFP C3 clone (a) and HT-29-NF-κB-hrGFP E5 (b) and F6 ((c), (d)) clones simultaneously with TNF-α ((a), (b), and (c)) or LPS (d), respectively. For HT-29-NF-κB-hrGFP clones, 1 ng/mL TNF-α or 5 ng/mL LPS was employed, while for Caco-2-NF-κB-hrGFP C3 clone 5 ng/mL TNF-α was used. After 48 h for Caco-2-NF-κB-hrGFP clone and 18–24 h for HT-29-NF-κB-hrGFP clones, NF-κB activation (measured by the percentage of GFP⁺ cells) was analyzed by flow cytometry. Cell viability was over 90% for all tested conditions. Data was normalized against TNF-α or LPS controls (considered as 100%) and shown as the mean ± SD of triplicates of a representative experiment. using One-Way ANOVA with Dunnett’s posttest.

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