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

Figure 4

NF-κB activation stability with passage number for different clones. Cells with 0 (black circle), 4 (downward triangle), or 8 (upward triangle) splitting passages of the reporter cell lines were seeded as follows: Caco-2-NF-κB-hrGFP C3 clone ((a) and (b)) at 1 × 10⁵ cell/well and HT-29-NF-κB-hrGFP E5 clone ((c) and (d)) or F6 clone ((e) and (f)) at 5 × 10⁴ cell/well. Cells were cultured ON and treated with TNF-α, LPS, or IL-1β. Cells were trypsinized and GFP expression was analyzed by flow cytometry. Data was normalized and fitted using nonlinear regression 3-parameter fit using GraphPad Prism. NF-κB activation was determined as previously described.

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