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Fig. 7. Generation of the Map2k1 conditional allele. (A)
Map2k1 gene targeting strategy for the generation of the conditional
allele. Map2k1 translated exons 2 to 4 are represented by the white
boxes. A neo selection cassette flanked by loxP sites was
inserted between the third and fourth Map2k1 exons. A third
loxP site was inserted between the second and the third exons. The
targeting vector contains 5.5 kb and 6.9 kb of Map2k1 homologous
genomic sequences on the 5' and 3' sides of the neo
insertion, respectively. The herpes simplex virus-thymidine kinase (HSV-TK)
selection cassette (gray box) was added at the 5' end of the vector.
(B) Southern blot analysis of tail DNA from a litter obtained after
breeding a Map2k1+/floxed female with a
Sox2Cre+/-;Map2k1+/
male. Tail
DNA was digested with StuI, blotted and hybridized with a
Map2k1 genomic probe (A; upper panel) and a Cre probe (lower
panel). The position of the different alleles is indicated on the right side
of the panels. (C) Western blot analyses of proteins extracted from
different organs of 8-week-old wild-type (+/+) and
Map2k1/ (/) littermates.
Triplicate blots were probed with antibodies directed against MAP2K1, MAPK1
and MAP2K2. Br, brain; He, heart; Ki, kidney; Li, liver; Lu, lung; Pa,
pancreas; Th, thymus.
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