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Fig. 2. Cre-mediated recombination of the conditional allele results in loss of
cilia and Ift88 protein. (A-C) Cilia were identified in frozen
sections of E11.5 wild-type (A) and prx1cre;Ift88fl/n
conditional mutant (B,C) limbs by immunofluorescence staining with antibodies
to detect acetylated 
-tubulin (green) and Ift88 (red). Nuclei are shown
in blue. In wild-type samples (A), cilia were visible on most cells in the
mesenchyme. By contrast, cilia were rarely observed on cells in the mesenchyme
of conditional mutants (B). The presence of cilia on the ectoderm of
prx1cre;Ift88fl/n mutant limbs (C) was not affected.
(D) Protein lysate from conditional mutant limbs at E11.5 (prx1cre)
showed a dramatic reduction in the amount of Ift88 expressed when compared
with lysate from a control littermate (WT). Samples were simultaneously
blotted for ß-tubulin as a loading control. Approximate molecular weights
are shown to the left of the blot. (E,F) At E11.5, cilia are
apparent in the dorsal ectoderm of the hindlimb in
msx2cre;Ift88fl/n mice (E) by immunofluorescence staining
for acetylated -tubulin (green) and Ift88 (red), but are absent from
the ventral ectoderm of the same limb (F).
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