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

Table 2

NF-B activation stability with passage number for different clones.

Passage number	Caco-2-NF-B-hrGFP		HT-29-NF-B-hrGFP
	Clone C3		Clone E5		Clone F6
	TNF-	IL-1	TNF-	IL-1	TNF-	LPS
0	ND	0.23 (0.16–0.33)	0.52 (0.47–0.59)	0.21 (0.15–0.28)	0.72 (0.43–1.22)	0.15 (0.10–0.21)
4	ND	0.12 (0.09–0.16)	1.03 (0.77–1.38)	0.47 (0.30–0.75)	2.80 (2.58–3.08)	0.45 (0.32–0.63)
8	ND	0.20 (0.12–0.33)	1.00 (0.86–1.16)	0.44 (0.23–0.81)	4.30 (3.72–0.81)	0.33 (0.24–0.44)

Different cell splitting passages of the reporter cell lines were seeded. 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 EC50 (ng/mL) was estimated using nonlinear regression 3-parameter fit using GraphPad Prism. 95% confidence intervals are shown between brackets. ND: not determined.