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Fig. 5. Ecdysone signalling cell autonomously controls all aspects of cardiac tube
remodelling. In all cases, individuals were shifted to 29°C at the onset
of metamorphosis. (A) Phalloidin staining of a Gal80ts,
24B>Gal4; UAS>EcRDN pharate adult cardiac tube. Inhibition of
EcR function prevents heart remodelling, including A6/A7 histolysis (red
asterisk), A5 transformation (arrowhead) and posterior aorta remodelling,
which remains thin anterior to A5 (arrow). (B) Gal80ts,
24B>Gal4; UAS>dsRNA-EcR cardiac tube at 30 hours APF, stained
for Wg, detected with anti-mouse alkaline phosphatase-conjugated secondary
antibody. wg is not expressed in the svp-expressing myocytes
(arrowheads). (C) Phalloidin (white) and anti-Ubx (red) staining of
NP5169-Gal4; UAS>dsRNA-EcR pharate adult cardiac tubes. EcR inactivation in
Tin-expressing myocytes prevents Ubx repression in segment A1-A4
Tin-expressing myocytes (arrows). pc, pericardial cell. (D) Anti-synaptotagmin
(green), 22C10 (red) and phaloidin staining of Gal80ts,
24B>Gal4; UAS>EcrDN pharate adult cardiac tubes. (E) Anti-AbdA (green)
and phalloidin staining on Gal80ts, 24B>Gal4;
UAS>EcRDN pharate adult cardiac tubes. In both cases, myofibrils
are transversal (open arrow), indicating that EcR loss of function prevents
terminal chamber differentiation. (D) Nerve terminations do not make synaptic
contact with the myocytes, as judged by the absence of synaptotagmin staining.
(E) EcR inactivation does not affect AbdA expression. (F,G) Phaloidin staining
of Gal80ts, 24B>Gal4; UAS>AbdA, UAS>EcRDN
pharate adult cardiac tubes. Inhibition of ecdysone signalling prevents AbdA
from inducing terminal chamber structure both in A1-A4 segments (F) and in A5
(G). (H) Schematic representation of a cardiac tube in which EcR function has
been inhibited during metamorphosis. Ubx and abdA expression are
represented (see C and E). Scale bars: 50 µm.
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