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Supplementary material for this article can be found on the H EPATOLOGY website ( http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html ).

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jws-hep.21528.fig1.tif14568KSupplementary Fig. 1.Full scheme of bile salt synthesis and organelles involved in the liver.Bile salts are synthesized in the hepatocytes in a series of reactions that involves several cell compartments. The key regulating enzyme of the classic or neutral pathway?starting in the ER?is CYP7A1. The first step of the alternative or acidic is CYP27A1, located in the mitochondrion. The two pathways converge in the peroxisome where β-oxidation and glycine/taurine conjugation of the side chain occur. Conjugated bile salts are substrates for BSEP for transport to the bile. The fraction of reabsorbed bile salts that are de/un-conjugated require reactivation with CoA, which is catalyzed by FATP5. Subsequently, Co-A-activated bile salts require transport into peroxisomes for glycine or taurine conjugation by BAAT. Next, the reconjugated bile salts, together with thede novosynthesized conjugated bile salts in peroxisomes require export out of this organelle before they can be secreted to the bile by BSEP.
jws-hep.21528.fig2.tif15327KSupplementary Fig. 2.Immunofluorescence microscopy for hBAAT in human hepatocytes reveals a typical peroxisomal staining.Human hepatocytes were allowed to attach to coverslips and immediately processed for immunofluorescence microscopy with antibodies against hBAAT as detailed in Material and Methods. Pictures were captured as indicated in the Materials and Methods section. A sequence of 25 pictures were taken, at focal distances 0.499 μm apart. The same dotted peroxisome-like pattern is observed throughout the cell.
jws-hep.21528.tbl1.pdf22KSupplementary Table 1.Primers and probes used for Q-PCR
jws-hep.21528.tbl2.pdf22KSupplementary Table 2.Cloning primer sets used in this study

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