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Fig. 8. Zebrafish vps33b promoter is regulated and bound by Hnf1. (A) Sequence of the putative zebrafish promoter. Predicted Hnf6 binding sites are highlighted in yellow, predicted vHnf1 binding sites are underlined and in blue, and the predicted sequence of the first untranslated exon is noted in green. (B) Expression of a luciferase vps33b reporter gene [vps33b(-1560/+139)-luciferase] is activated in BMEL cells by vHnf1, but not Hnf6 or GFP proteins. ^*P<0.05. (C) Electrophoretic mobility shift assays performed using nuclear extracts from BMEL cells that were transfected with GFP (lane 1) or vHnf1 (lanes 2-5) expression constructs. Lanes in which the extracts are incubated with anti-vHnf1 antibody (vHnf1; lane 3), anti-Hnf-1 antibody (Hnf-1; lane 4), or 50x excess cold probe directed against the vHnf1-4 site (lane 5) are noted. Addition of cold probe eliminates supershift seen in lane 3. Arrows indicates the gel shift resulting from vHnf1 binding (bound vHnf1) or vHnf1/anti-vHnf-1 antibody binding (supershifted vHnf1) to the labeled probe.

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