Xenbase Image ID: 26055

Fig. 5. Expression of the 152 bp distal enhancer is dependent upon conserved Pax6 sites and Pax6 binds to this region in vivo. (A) Sequence from the Xenopus laevis 152 bp enhancer region showing the 2 overlapping Pax6 sites identified using Transfac MATCH version 10.3 (http://www.gene-regulation.com/pub/databases.html#transfac). P1 aligns with the Pax6_01 matrix, while P2 aligns with the Pax6_Q2 matrix. (B) Mutation of one Pax6 site dramatically reduces (−/+) while mutation of both Pax6 sites in the 152 bp distal enhancer results in retinal GFP expression in only a few cells (−/(+)) in transgenic embryos. (C) A stage 33 pG1X5 distal 152 bp transgenic embryo shows strong GFP expression in the retina. (D) A stage 33 pG1X5 distal 152 bp P1P2 mut transgenic embryo does not have retinal GFP expression. (E) ChIP showing Pax6 binds to the distal enhancer region in vivo (group student t test P = 0.002 for enhancer region (IgG vs. Pax6 antibody), and 0.056 for the GFP coding region (IgG vs. Pax6 antibody)). Error bars represent standard deviation. Gut autofluorescence. Image published in: Willardsen MI et al. (2009) Copyright © 2009. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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