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Fig. 1. A Mef2c-lacZ bacterial artificial chromosome (BAC) transgene is
sufficient to direct the complete pattern of Mef2c expression. (A)
Schematic representation of the mouse Mef2c locus contained within
BAC clone GS133. The BAC contains the first six known exons from
Mef2c (vertical black lines), including the first two coding exons,
which encode the MADS and MEF2 domains of Mef2c. The red box
represents a region of high evolutionary conservation that functions as an
anterior heart field specific transcriptional enhancer. (B) Schematic
representation of Mef2c-F6-lacZ, which contains the Mef2c
anterior heart field enhancer subcloned into the transgenic reporter plasmid
HSP68-lacZ. Red box, Mef2c noncoding sequences representing
the anterior heart field enhancer; black box, Mef2c untranslated exon
3; green box, HSP68 promoter (pr); blue box, lacZ gene; yellow box,
SV40 splice and polyadenylation (polyA) sequence. (C) Expression of the
Mef2c BAC transgene, Mef2c-L8-lacZ, at 7.5 days
post coitum (dpc) shown in frontal view. The Mef2c-L8-lacZ
BAC transgene directs expression exclusively to the anterior heart field at
this stage, recapitulating the earliest expression of endogenous
Mef2c. (D) Lateral view of a transgenic embryo collected at 8.0 dpc
showing expression of Mef2c-L8-lacZ. Strong expression is
evident in the anterior heart field at the arterial pole of the linear heart
tube. (E) Ventral view of an 8.5 dpc Mef2c-L8-lacZ
transgenic embryo. Strong expression remains evident in the anterior heart
field at the arterial pole of the linear heart tube. Transgene expression is
also evident in the neural crest (NC) at this stage and expression can begin
to be seen in vascular endothelial cells within the dorsal aortae and in the
yolk sac (not shown). (F) Lateral view of a 9.5 dpc
Mef2c-L8-lacZ transgenic embryo. At this stage the
expression directed by the BAC transgene completely recapitulated the pattern
of endogenous Mef2c expression during embryonic development at this
stage. Expression was present in vascular endothelium (V), neural crest,
somites (S), branchial arches (BA), pharyngeal mesoderm, outflow tract and
heart. RV, right ventricle. (C-F) Representative X-gal stained transgenic
embryos at each stage. Arrowheads point to pharyngeal mesoderm/anterior heart
field. Two F0 transgenic embryos and embryos collected from an
independent BAC transgenic line displayed identical patterns of expression at
all stages.