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Fig. 4. Deletion of Myog and attenuation of myogenin transcripts and
protein in Mygflox
/flox mice. (A)
Quantitative PCR of tail DNA to determine the extent of deletion of floxed
Myog sequences in Myogflox/flox and
Myogflox/flox mice at P10. The ratio of
Myog to Myf5 genomic DNA from one
Myogflox/flox mouse was arbitrarily set to 1 and all other
ratios from both Myogflox/flox and
Myogflox/flox genotypes were normalized to
that value. (B) Southern genome hybridization with
EcoRI-digested DNA extracted from the hindlimb skeletal muscle of
10-week-old mice from a Myogflox/flox;+/+ x
Myogflox/flox;CAGGCre-ERTM/+ cross and probed with
Myog cDNA. The 6 kb and 3 kb bands represent the
Myogflox and Myogflox alleles,
respectively, as depicted in Fig.
1. The skeletal DNA in lanes 1 and 3 contained very little of the
Myogflox allele and therefore represents a
Myogflox/flox genotype, while the skeletal
DNA in lanes 2, 4, 5 and 6 contained no Myogflox
allele and therefore represents a Myogflox/flox genotype.
(C) Quantitative RT-PCR to determine the levels of Myog
transcript expression in hindlimb skeletal muscle from mice at 3 days (right
panel) and 2 weeks (left panel) of age. The genotypes are shown below the
histograms. The ratio of Myog to ribosomal protein L7 RNA for one RNA
sample with a Myogflox/flox genotype was arbitrarily set
to 1, and all other ratios from both Myogflox/flox and
Myogflox/flox genotypes were normalized to
that value. The median value for each genotype is shown as a black dot.
Asterisks indicate significant differences between the two genotypes. The
arrow bars indicate one s.d. from the median. (D) Hindlimb sections
from Myogflox/flox (right panel) and
Myogflox/flox (left panel) mice at 3 days of
age immunostained with the monoclonal F5D anti-myogenin antibody. Arrows
indicate positively stained cells. Scale bar: 100 µm.
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