Figure 3.

LPS-induced LCN2 is dependent on NF-κB activation in macrophages. (A) After stimulation with 50 ng/ml LPS for the indicated time, cytosolic and nuclear extracts of RAW264.7 cells were prepared and used to determine activation of the NF-ĸB pathway. Actin and lamin B were used as loading controls. (B) Cells were pretreated with PT and BAY 11-7082 (NF-κB-specific inhibitor), and the cells were stimulated with LPS for 24 h. Then, the levels of LCN2 mRNA and protein were detected using RT-PCR (upper panel) and western blot analysis (lower panel). GAPDH was used as a loading control. LPS, lipopolysaccharide; LCN2, lipocalin 2; PT, parthenolide; RT-PCR, reverse transcription-PCR; IκB-α, NF-kappa-B inhibitor α.