Figure 4
IκBζ is induced by MALP-2 and regulates Lcn2 promoter activity. (A) HC11 cells were treated with 10 µg/ml LPS or 10 ng/ml MALP-2. IκBζ mRNA was quantified by RT-qPCR and normalized to the cyclophilin mRNA in the same sample and then all values were normalized to the 0-time point. (B) HC11 cells were cotransfected with the 253 bp Lcn2-luciferase plasmid and the Renilla-luciferase control plasmid in combination with a plasmid to express one of the following RNAs: SHC002 or shIκBζ. The transfected cells were treated with 10 ng/ml MALP-2 for 18 h then lysates tested for promoter activity. (C) HC11 cells were cotransfected with expression vectors for eGFP with the following combinations: shIκBζ or the SHC002, each in combination with IκBζ or mIκBζ. Samples were collected 24 h after transfection and exogenous IκBζ mRNA was quantified by RT-qPCR with primers that did not amplify the endogenous IκBζ. Each value was normalized to the level of eGFP mRNA in that sample.
