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Fig. 2. Identification of the functional E-box motifs responsible for the
somite-specific activation of Epha4. (A) Sequence of the
630 bp Epha4 somite enhancer (core enhancer) region. The four
indicated fragments (fragment 1-4) represent deleted sequences from the core
enhancer region. The results of transgenic analyses using these deleted
constructs are shown in parentheses. (B) Sequence alignment of somite
enhancer regions (fragment 2 and fragment 3) of mouse Epha4 with the
corresponding regions of human Epha4. The mutations introduced in
each E-box and the results of the subsequent transgenic analyses are shown.
(C) Schematic representation of artificially constructed enhancers,
containing six E-box motif repeats. (D) The artificial enhancers were
cloned upstream of a lacZ reporter vector and the results of the
subsequent transgenic analyses (representative images are shown) are indicated
in parentheses. For the [E2]6-lacZ artificial enhancer, no somite
expression was observed among the 11 transgene-positive embryos. Blue letters
indicate putative E-box motifs.
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