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. 2008 Oct 17;283(42):28095–28105. doi: 10.1074/jbc.M708350200

FIGURE 7.

FIGURE 7.

Hypertonicity-induced NF-κB activation is dependent on IKKβ activity, IκBα degradation, and p65 activation. A and B, cells were transfected with an expression vector that contained either eGFP or constitutively active IKKβ or IκBα mutants together with (A) or without (B) NF-κB-driven luciferase plasmid. Cells were challenged or not with either 100 μg/ml LPS or hypertonic (NaCl, 500 mosmol/kg) medium for 12 (A) or 3 (B) h before measurement of luciferase activity (A) or to RNA extraction (B). Real-time PCR was performed using primers specific for TNFα (B). Results are expressed relative to control values determined in cells transfected with eGFP-containing cDNA and subjected to isotonic medium. Bars are the mean ± S.E. from four independent experiments. *, p < 0.05. C, cells were transfected with either scrambled RNAi or RNAi targeting p65 NF-κB and then challenged or not with either 100 ng/ml LPS or hypertonic (NaCl, 500 mosmol/kg) medium for 3 h before RNA extraction. Real-time PCR was performed using primers specific for MCP-1, IκBα, or TNFα. Results are expressed relative to control values determined in cells transfected with scrambled RNAi and subjected to isotonic medium. Bars are the mean ± S.E. from four independent experiments. *, p < 0.05. Ctl, control.