Manuscript number: C0517a
Title: Distalization of the Drosophila leg by graded EGF-receptor activity
Author: Gerard Campbell
Legends for the Supplementary Figures
Supplementary Figure 1. Companion to main text Figure 1. a-b, Single channel
images of Fig. 1b. c, Merged image of Fig. 1c, d showing clear overlap between
rn and dac (yellow). d, Late third instar leg disc from a wgts mutant raised at the
restrictive temperature during the third instar (i.e. as for the disc shown in Fig. 1f)
and stained for Dll expression. Dll is still expressed in the center of the disc (Dll
expression is only dependent upon WG during the second instar). This
demonstrates that loss of al following loss of WG at the beginning of the third
instar is not the result of loss of Dll. e, f, h, Single channel images of Fig. 1h. g,
An additional channel from the same disc showing Dll expression, which, as in d,
is still expressed in the center in arr mutant cells (arrow), so again the loss of al is
independent of Dll. i-k, Single channel images of Fig. 1i. l-n, Single channel
images of Fig. 1j.
Supplementary Figure 2. Companion to main text Figure 2. a-c, Adult legs from
Egfrts mutants raised at restrictive temperatures for the first half of the third
instar. a, At 31oC the only defect is seen in the tarsus, the rest of the leg being
normal. b-c, At 33oC many legs also only have defects in the tarsus (b), while
others (c) show defects in more proximal regions. d, If the shift is made during
the second instar the legs are patterned normally. e-f, Single channel images of
Fig. 2d. g-h, Single channel images of Fig. 2e. i-j, Single channel images of Fig.
2f. k-l, Single channel images of Fig. 2g. m, Wild-type disc stained for dpp
expression alongside the disc shown in (n) and Fig. 2h. o, Wild-type disc stained
for wg expression alongside the disc shown in (p) and Fig. 2i. q-t, Single
channel images of Fig. 2j. u-w, Single channel images of Fig. 2k. x-z, Single
channel images of Fig. 2l.
Supplementary Figure 3. Companion to main text Fig 3. a, vn null mutant leg
disc showing normal al expression in the center. b, Adult tarsus comprised
exclusively of rho mutant clones. This is patterned normally. Similar legs
(particularly metathoracic legs) may show abnormal joint formation between the
tarsus and the tibia (not shown). c-d, Tip of a leg containing ru rho mutant cells.
One of the claws is wild-type and appears normal (c), while the other (d) is very
rudimentary with no pigmentation (even y claws have some melanization). This
is an extreme example, others are usually just smaller with no pigmentation.
This phenotype is never seen with rho single mutant clones. ru single mutant
adults are viable with completely normal legs. spitz mutant clones have no effect
on leg patterning, even the claw (not shown), suggesting RU and RHO are
activating another TGF-alpha, such as Keren (no mutations are currently
available in the gene encoding for this).