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Fig. 3. Prolongation of the transient enhancer activity of the ASE of
Lefty2 by addition of Nkx2-binding sequences. (A,B)
Structures of two lacZ transgenes are shown at the top. Red circles
and blue ovals indicate Foxh1- and Nkx2-binding sequences, respectively. The
partially blue circles and boxes indicate the hsp68 promoter and
lacZ, respectively. The construct in A includes the Lefty2
ASE, which contains two Foxh1-binding sequences. The construct in B contains
the Lefty2 ASE plus seven tandem repeats of the Nkx2-binding sequence
derived from the Pitx2 ASE. X-gal staining patterns of E8.2 mouse
embryos harboring each transgene are shown below. Both lacZ
constructs gave rise to typical asymmetric staining in left LPM. (C-H)
Transverse sections of X-gal-stained transgenic embryos at E10.5. Expression
of lacZ is shown for the common atrial chamber (C,D), foregut (E,F),
and gut dorsal mesentery (G,H). Black arrows indicate the domains where
asymmetric X-gal staining was detected. Encircled + or - signs indicate
whether the enhancer was active or inactive, respectively. The Lefty2
ASE construct did not give rise to asymmetric staining (C,E,G), whereas
the N7-Lefty2 ASE construct did so in the common cardinal
vein, lung bud and gut dorsal mesentery (D,F,H). Scale bars: 200 µm.
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