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Fig. S3
Homologs of ethanol metabolizing enzymes are expressed in zebrafish. (A) The protein sequence alignment of zebrafish (Danio rerio; Dr) Cyp2y3 and Cyp2p6 with that of human CYP2E1 (Hs) and mouse Cyp2e1 (Mm). Cyp2y3 and Cyp2p6 show 43% identity to human CYP2E1. The pink box highlights the substrate-binding site, which is also a part of the antibody-binding site. (B) Representative PCR agarose gels of adh5, adh8a, adh8b, cyp2y3, cyp2p6, aldh2 performed on cDNA prepared from a pool of dissected livers from control (–) and larvae treated with 350 mM ethanol (+) for 32 hours. rpp0 serves as a reference gene. (C-E) Immunoblots of proteins from 10-25 livers per sample using antibodies raised against human ADH1 and CYP2E1. VL-17A cells stably expressing mouse Adh1 and human CYP2E1 served as a positive control.