Iron deficiency induces a proinflammatory condition. (A-B) Analysis of liver STAT3 activation. Livers were dissociated as described and protein extracts were subjected to SDS-PAGE and Western blotted for phospho-STAT3 and STAT3 to normalize gel loading. A representative Western blot is shown. (C) The level of STAT3 phosphorylation was quantified by densitometric analysis of phospho-STAT3 normalized to STAT3. Hepatic expression of the inflammatory cytokines IL6 (D) and TNFα (E), measured by SybrGreen qRT-PCR, and of acute-phase proteins A2M (F) and serum amyloid A-1 (SAA-1; G), measured by TaqMan qRT-PCR (10 mice each group). Gapdh (SybrGreen qRT-PCR) or Hprt1 (TaqMan qRT-PCR) were used as housekeeping genes to normalize gene expression. mRNA expression ratio was normalized to an IR mean value of 1. a.u. indicates arbitrary unit. Error bars indicate SE; ns, not significant; *P < .05; **P < .005; and ***P < .0001.