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Fig. 1. RNAi silencing of dally-like but not dally generates a
segment polarity phenotype in Drosophila embryos. (A) Ventral view of
a wild-type larval cuticle. Unless specified, the head of embryos is to the
left. Eight belts of denticles are visible in the abdomen. (B and C) Segment
polarity phenotype of null mutations in wingless
(wgCX4) and hedgehog (hhAC).
The areas of naked cuticle are replaced by denticles, generating a lawn of
denticles without clear polarity. (D) Position of the dsRNA sequences (in red)
and the RT-PCR sequences (in blue) used in dally and
dally-like RNAi experiments. (E) Agarose gel electrophoresis of
semiquantitative RT-PCR reactions performed on extracts of embryos injected by
either buffer, dlp 3'dsRNA or dally 3'dsRNA.
There is at least a fourfold decrease in dlp mRNA following
dlp RNAi, and at least a fivefold decrease in dally mRNA
following dally RNAi. By contrast mRNA levels of dally and
dlp following dlp and dally RNAi, respectively, are
identical to the levels after injection of buffer. (F) Weak segmentation
defects exhibited by dally dsRNA-injected embryos. The same type of
defects are found in buffer-injected embryos (not shown), and thus are a
consequence of the injection process. There is no clear transformation of
naked cuticle into denticles in these embryos, but rather a loss of tissue
that leads to the fusion of denticle belts. (G-I) Segment polarity defects
found in dlp dsRNA injected embryos. (G) The weak segment polarity
phenotypes are distinct from the phenotypes observed in buffer and
dally-injected embryos: the naked cuticle is transformed into
denticles without much tissue loss. (H) Strong segment polarity phenotype and
(I) lawn of denticles identical to the phenotype of wingless or
hedgehog null mutants. (See also
Table 1.)
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