Fig. 3. Characterization of zebrafish ABCB6 (Abcb6).

a Alignment of homologous ABCB6 sequences shows zebrafish Abcb6 lacks the N-linked glycosylation site⁸⁹. b Western blot of samples from PNGase F treatment confirms that zebrafish Abcb6 is not a glycoprotein, unlike its human homolog (N = 1 as experiment is a qualitative confirmation of sequencing data). Western blots show zebrafish Abcb6 binds ATP-agarose (c) and various porphyrins like its human homolog (d). Representative data shown, N = 3 biological replicates for each pulldown, with each experiment repeated from transfection to western blot. e Overlay of a representative structure from the MD simulations of the zebrafish Abcb6 (green) and ABCB6 WT (indigo/turquoise as shown in Fig. 2) showing that the two proteins adopt a similar conformation. f Overlay of zebrafish Abcb6 WT and zebrafish Abcb6 L356P showing the change in conformation of the coupling helix. Source data are provided in the Source Data file.