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Fig. 7. Shh signaling and cholesterol synthesis. (A) Various DHCR7
mutant constructs. (B) Inhibition Hh signaling is enhanced by treatment
with AY-9944. Embryos were injected with the reporter construct
8X3'Gli-BS Luc with or without Shh-N, DHCR7 mRNA and treatment
with 10 µM AY-9944. (C-J) Microinjection of DHCR7 mutants that are
defective in reductase activity causes cyclopic phenotypes. The overexpression
of DHCR7 mRNA (E) or treatment with AY-9944 (D) caused minor defects,
but a combination of both causes severe defects (F). Embryonic phenotypes
caused by microinjection of 1 ng of DHCR7R350W (H),
DHCR7IVS8-1G>C (G),
DHCR7
N (J) and
DHCR7W149X (I) mutants. (K) Pax6 staining
of tailbud stage embryos injected with DHCR7R350W. Arrow marks
`cyclopic' eye. (L) Modulation of Gli reporter by DHCR7 mutants using
animal cap conjugation. Embryos were injected with Chordin (with or
without Shh-N mRNA) or with Gli-BS Luc (with or without
DHCR7 mRNA).