TLR4 dependency of bacterial-induced hepcidin mRNA expression by neutrophils and macrophages. (A) Real-time PCR for hepcidin mRNA on WT (CH3/HeouJ) and Tlr4^Lps-d (CH3/HeJ) murine macrophages stimulated with P aeruginosa (PA), S typhimurium (ST), LPS, or Group A Streptococcus (GAS) producing or lacking the cytotoxin streptolysin S (SLS). Hepcidin mRNA was normalized to the expression level of β-actin. Quantitative assays performed in triplicate and representative of 3 repeated experiments. (B) Real-time PCR for hepcidin-1 and hepcidin-2 mRNA in WT (CH3/HeouJ) and Tlr4^Lps-d (CH3/HeJ) macrophages stimulated with PA or LPS. Data are shown as mean of values ± standard deviation (SD).