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Fig. 2. Targeted disruption of the mouse Mll2 gene. (A) The
mouse Mll2 cDNA sequence is illustrated at the top to indicate the
positions of domains and motifs (see Fig. S1 in the supplementary material for
more details). Below is a diagram of the gene with exons shown as numbered
boxes connected by known splicing events. The exact start site of
transcription is not known but arises in the indicated CpG island.
Polyadenylation signals are indicated by red circles. The RT-PCR primers and
in situ probe are indicated below, as are the alleles after targeting, FLPe
and Cre recombination. (B) RT-PCR analysis of E8.5 embryos. An
Mll2-specific primer pair amplified a reaction product from
Mll2+/– and Mll2+/+ embryos, but
not from a Mll2–/– embryo. (C) Western
blot analysis of wild-type and Mll2–/– ES
cells probed with an antibody raised against Mll2 amino acids 864-980. The
strong band detected in wild-type extract probably corresponds to the
predicted proteolytic 225 kDa Mll2 fragment processed by Taspase. The weaker
band in wild-type extract could correspond to the 284 kDa full-length protein.
No bands were detected in extracts from
Mll2–/– cells and the blot was controlled
using antibodies against CBP, which is also very large. (D) Expression
of Mll2 at E8.5. The expression is ubiquitous in the wild-type embryo
and absent in the homozygous embryo. (E,F)
Mll2–/– embryos and Mll2FC/FC embryos
have an identical embryonic lethal phenotype, as illustrated here for
E9.5.
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