Kinetic analysis of LPS-induced inflammatory responses in wild-type mice. Five-week-old male wild-type C57BL/6 mice (n = 6 for each experimental group) were left untreated or injected with 1 μg/g LPS and sacrificed after 2, 4, 8, 18, or 24 hours. Sera were used for iron biochemistry, and tissues were processed for preparation of RNA and protein extracts or fixed for histology. (A) qPCR analysis of liver Il6 mRNA; (B) western blot analysis of liver pStat3 and Stat3; (C) qPCR analysis of liver ferroportin (Fpn + IRE) mRNA (IRE-containing isoform); (D) qPCR analysis of liver Hamp mRNA; (E) serum hepcidin; (F) serum ferritin; (G) serum iron; (H) transferrin saturation; (I) TIBC. All data in the graphs (A-I) are presented as the mean plus or minus standard error of the mean (SEM). Dotted lines indicate average values obtained from age-matched male C57BL/6 mice (n = 3) fed a high-iron diet for 5 days. (J) Immunohistochemical staining of ferroportin in liver and spleen sections (original magnification ×20 and ×40). Arrows indicate ferroportin internalization in Kupffer cells.