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. 2015 Jan 13;5:7750. doi: 10.1038/srep07750

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(a) In Xenopus Hnf1b induces mafb and krox20 indirectly through the activation of egr4. (b) In the mouse Hnf1b directly activates mafb and krox20 expression. (c) The expansion of mafb in embryos injected with hnf1bGR mRNA (10 pg) is completely blocked by co-injection of egr4MO2, but not by a control morpholino (ctrMO). Scale bar, 300 μm. The graph indicates the percentage of embryos with normal (white), reduced/lost (red) or expanded (blue) mafb expression. The number of embryos analyzed is indicated on top of each bar. (d) A conserved region upstream of the Mafb gene containing and Hnf1 binding site has been characterized as an r5/r6-specific enhancer (S5). (f) A putative S5 enhancer (pS5) downstream of Xenopus tropicalis egr4 containing two putative Hnf1 binding sites was identified using rVISTA 2.0. (e, g) Schematic representation of the S5 and pS5 enhancer around Mafb (mouse) and egr4 (Xenopus) genes, respectively. (h) Experimental procedure to test the activity of the pS5 in Xenopus embryo. (i) Fold EGFP induction analyzed by qPCR. Values are normalized to ef1α. Graph represents mean ± S.E. of 3 independent experiments. *, P<0.05; **, P<0.01; versus embryos injected with each vector alone. Student's t-test.