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Fig. 1. An Xvent2-BRE reporter mimics brk expression in
Drosophila and assembles a Mad/Med/Shn protein-DNA complex.
(A) Reporter constructs. (B-Q) Top row, lateral views of stage
13 embryos; anterior left, dorsal up. Succeeding rows show third-instar wing,
eye-antennal and leg imaginal discs stained to visualize lacZ
expression. Grh-lacZ drives nearly ubiquitous expression during (B)
late embryogenesis, and (C-E) in imaginal discs. (F-I) The
Xvent2-BRE-lacZ reporter is Bmp sensitive. lacZ
expression is downregulated at sites of high Dpp signaling. (J-M) Expression
of a brklacZ reporter closely matches expression of
Xvent2-BRE-lacZ. (N) In situ hybridization showing sites of
dpp expression in the embryo. (O-Q) dpp-lacZ expression in
imaginal discs. The leading edge of the dorsal ectoderm is marked with a bar
in F and J, and an arrow in N; arrowhead in N indicates the boundary between
the dorsal and ventral ectoderm. (R) Lysates from S2 cells transfected
with the indicated plasmids were used to gel shift the BRE probe. The presence
of Mad/Med results in a low mobility complex (band a, lane 2) that is further
retarded by anti-FLAG (band b, lane 3) or Shn-Myc (band c, lane 4). The latter
complex is supershifted by incubation with anti-Myc (band d, lane 5).
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