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. 2011 Dec 22;6(12):e29183. doi: 10.1371/journal.pone.0029183

Figure 2. Ablation of hepatic Commd1 expression results in increased intracellular copper concentrations accompanied with decreased Atp7b expression in young mice.

Figure 2

A.) Hepatic copper concentrations were measured in dried liver tissue of Commd1 loxP/loxP (white bars; n = 6) and Commd1 Δhep mice (black bars; n = 5) (6, 9 and 12 weeks of age) by means of FAAS. Data are represented as the hepatic copper concentrations (µg/g dry liver weight) in all mice at indicated ages. * indicates significantly different values compared to Commd1 loxP/loxP mice (p<0.05). B.) Relative mRNA expression of metallothioneins Mt-I and Mt-II in liver tissue of Commd1 loxP/loxP (open dots; n = 6) and Commd1 Δhep mice (black dots; n = 5) (six weeks of age) as determined by qPCR analysis. Expression was normalized for β-Actin mRNA levels, and relatively expressed to Commd1 loxP/loxP mice. C.) Immunoblot analysis of liver tissue of Commd1 loxP/loxP and Commd1 Δhep mice at six weeks of age. 30 µg of liver homogenates were analyzed by SDS-PAGE, and immunoblotted (IB) for expression of Atp7b and Commd1. Actin protein expression was used as the loading control. D.) Densitometric quantification of Atp7b expression at six weeks of age (Figure 2C), normalized to Actin expression. Mean expression of Commd1 loxP/loxP mice was set at 1 ± SD. p = 0.0010. E.) Relative mRNA expression of Atp7b in liver tissue of Commd1 loxP/loxP (open dots; n = 5–6) and Commd1 Δhep mice (black dots; n = 5–6) (6, 9 and 12 weeks of age) as determined by qPCR analysis. Expression was normalized for β-Actin mRNA levels, and relatively expressed to Commd1 loxP/loxP mice. F.) Immunoblot analysis of Atp7b expression in liver tissue of Commd1 loxP/loxP and Commd1 Δhep mice at 12 and 52 weeks of age.