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. 2012 Oct 24;7(10):e48679. doi: 10.1371/journal.pone.0048679

Figure 1. LPS differentially regulates ZIP14 expression in mice.

Figure 1

Young adult mice received LPS (2 mg/kg, i.p.) or the same volume (0.5 mL) of saline (control), 1–18 hr before being killed. (A) Total RNA was isolated and Zip14 mRNA was measured by qPCR and expressed relative to TBP mRNA as the normalizer. ZIP14 protein abundance was measured by western analysis of liver homogenates. Representative western blots from multiple mice (n = 3−4) were measured for ZIP14 abundance by densitometry. (B, C) Zinc concentrations in serum and liver, in µg/mL and µg/g respectively, were measured by AAS. (D, E) Comparison of Zip14 mRNA and ZIP14 protein in WT and Zip14 KO mice 18 hr after LPS, as measured by qPCR and western analysis. Values are mean ± SD, n = 3−5. (E) ZIP14 protein is increased at the plasma membrane of hepatocytes of WT mice but not Zip14−/− mice following LPS. Localization was by confocal microscopy using ZIP14 antibody and Alexa fluor594 secondary antibody and DAPI as the nuclear marker. (F) Serum IL-6 as measured by ELISA was used as an indicator of efficiency of LPS administration. The IL6 response from LPS was attenuated in the Zip14−/− mice. (* = P<0.05, ** = P<0.01, *** = P<0.001, **** = P<0.0001).