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Fig. 1 Subcellular localization and biochemical properties of zebrafish BCDO2. (A) COS7 cells expressing recombinant zebrafish BCDO2 with a V5 tag. Immunostaining was performed using anti-V5 antibody for BCDO2 (red) and mitochondria-specific anti-COXIV antibody (green). Merged image (right-most panel) shows mitochondrial colocalization of BCDO2 with COXIV. Images are taken at 63× magnification. (B) Recombinant zebrafish BCDO2 was expressed in an E. coli strain capable of synthesizing BC. Carotenoids and putative carotenoid-cleavage products were extracted and subjected to HPLC analysis. The solid line shows a HPLC profile at 420 nm of the E. coli lipid extract. Two major peaks (1 and 2) became detectable. These peaks showed similar retention times and spectral characteristics (insets) as the β-10′-apocarotenal oxime (syn and anti) standards (peaks 1′ and 2′) (dotted line). The insets give spectral characteristics of peak 1 and peak 1′.