CRBN elevates hepatic hepcidin gene expression in mouse primary hepatocytes. (A) Mouse primary hepatocytes were infected with Ad-GFP and Ad-Crbn at a multiplicity of infection of 60 for 36 hr. Gene expression was analyzed by qPCR using various primers. (B) Expression levels of various proteins in mouse primary hepatocytes were analyzed by immunoblotting using corresponding anti-bodies. (C) Expression levels of miR-204, miR-455, and miR-639 were quantified by qPCR analysis. (D) Mouse primary hepatocytes were infected with Ad-Scramble (Ad-Scram) and Ad-shCrbn at a multiplicity of infection of 60 for 36 hr. After infection, cells were treated with FSK (10 μM) for 12 hr. Total RNAs were utilized for qPCR analysis using gene-specific primers. (E) Whole-cell extracts were harvested from indicated groups of mouse primary hepatocytes and immunoblotted with specific antibodies. (F) WT mice were infected with Ad-Scram and Ad-shCrbn for 7 days. These mice were then fasted for 24 hr. Total RNAs were utilized for qPCR analysis using gene-specific primers. (G) Serum levels of hepcidin and iron at the indicated condition. *P < 0.05, **P < 0.01, and ***P < 0.001 vs. untreated control cells, Ad-GFP, or FSK-treated cells, fed, or fasted mice.