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Fig. 2. PDGFR
conditional allele and analysis of Cre efficiency. (A) The
wild-type PDGFR
locus, the targeted floxed allele and the resulting
allele after Cre-mediated inactivation of the PDGFR
. The deleted region
of genomic DNA in the fl allele is identical to that deleted in the
PDGFR
null allele
R4
(Soriano, 1997). Black boxes
represent exons. The floxed allele contains the neo cassette (gray box). Red
arrowheads represent loxP sites, and the small black rectangle represents the
probe used in Southern blot analysis. (B) Southern analysis of tissues from
E13.5 PDGFR
fl/fl; Wnt1Cre+ embryos. Genomic DNA
was digested with NheI and probed with fragment indicated in A. Lane
1, DNA from yolk sac; lane 2, DNA from cranial tissues. (C,D) Transverse
sections through first branchial arch region of E9 embryos stained for
ß-galactosidase activity. (C) ROSA26R+/-; Wnt1Cre+;
section was counterstained with nuclear Fast Red. (D)
PDGFR
fl/fl;Wnt1Cre+;ROSA26R+/-;
section was not counterstained. Note that D contains a similar distribution of
ß-galactosidase-expressing cells in the branchial arches when compared to
the distribution of ß-galactosidase-expressing cells in C. (E)
PDGFR
cell-surface expression histogram. Cells from E10.5 embryos were
isolated from the branchial arches, stained for PDGFR
expression, and
analyzed by flow cytometry. Red profile, cells stained with only secondary
antibody; gray profile, cells from PDGFR
fl/fl embryo; black
profile, cells from PDGFR
fl/fl;Wnt1Cre embryo.