SAA stimulates NF-κB activation and G-CSF transcript accumulation. (A) RT-PCR detection of G-CSF transcript in SAA-stimulated mouse BMDMs. β-Actin was used as a PCR and sample loading control. (B,C) Electrophoretic mobility shift assays showing SAA-induced binding of NF-κB (B) and CK-1 (C) to the respective DNA sequence in the promoter region of G-CSF, using nuclear extracts prepared from SAA- or buffer (CTL)–stimulated BMDMs. (D) A chromatin immunoprecipitation assay was conducted with SAA- or TNF-α (50 ng/mL)–stimulated RAW264.7 cells. An anti-p65/RelA antibody was used together with or without a specific blocking peptide. The immunoprecipitated DNA fragment was purified and amplified with PCR. DNA in total cell lysate was also amplified with PCR and used as a control. One representative experiment of a total of 3 is shown.