|
|
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Fig. 7. Vgll3 interacts with tead proteins and can activate pax3 independently
of tead. (A) Embryos injected with vgll3 (1ng or lacZ) mRNA and T1MO/T2MO or
cMO were fixed at stage 10 or 13 and analysed for pax3. Vgll3 induces ectopic pax3
expression when tead1 and tead2 have been knocked down. Lower right panel (IB
myc): embryos were injected with 50 pg of myc-tead1 or myc-tead2 mRNAs with 15
ng of T1MO or T2MO and analysed by IB. T1MO and T2MO block efficiently tead1
and tead2 expression respectively. Erk2 is used as control. ni: non injected embryo.
(B) Embryos injected with HA-vgll3, myc-tead1 or myc-tead2 mRNAs were processed for immunoprecipitation with HA (IP HA) or myc antibodies (IP myc)
followed by IB with antibodies. Vgll3 interacts with tead1 and tead2 in the embryo.
Lysate, IB of injected embryo before immunoprecipitation. Vertical white lines
indicate spliced images in order to remove results not described in this paper (IP
myc). |
|
|
|
Figure S8. The histidine repeat in vgll3 is required for pax3 activation. (A)
Sequence alignment of histidine (H) tract in Vgll3 proteins from different vertebrate
species. Numbers indicate the relative position of the histidine tract. Xenlae, Xenopus
laevis, Xentrop, Xenopus tropicalis. (B) Embryos were injected with 1 ng of vgll3 (vgll3
wt) or vgll3Dhis, fixed at stage 11 and analysed for pax3 expression. Vgll3Dhis cannot
activate pax3 (100%, n=30). (*) injected side. (C) HEK293 cells, recently tested for
contamination, were transfected with 1μg of plasmid encoding HA-vgll3wt or HAvgll3Dhis,
fixed and analysed by immunocytochemistry using HA antibody. The
deletion of histidine tract from vgll3 does not impair its nuclear localization. White arrow
indicates nuclear staining. Scale bars, 10 μm. |
|
Display additional annotations [+]
|
|
Figure S1. Vgll3 does not co-localise with trigeminal placode markers. (A) Wholemount
ISH for vgll3 and trigeminal placode genes islet1, neuroD, pax3 and foxi1c. (B)
Whole-mount ISH for trigeminal placodal genes islet1, neuroD, pax3 and foxi1c. Blue
arrow indicates the position of vgll3 expression domain. All staining are visualized in
blue. Analysed embryos are from stage 14 to stage 20 depending on the trigeminal
markers used. All views are dorsal-anterior. |
|
Gene |
Clone |
Species |
Stages |
Anatomy |
pax3.L
|
|
laevis
|
NF stage 15
|
neuroectoderm
,
neural crest
,
neural plate
,
preplacodal ectoderm
,
presumptive rhombomere
|
pax3.L
|
|
laevis
|
NF stage 18
|
neuroectoderm
,
neural crest
,
neural plate
,
preplacodal ectoderm
,
presumptive rhombomere
|
pax3.L
|
|
laevis
|
NF stage 19
|
neuroectoderm
,
neural plate
,
cranial neural crest
,
preplacodal ectoderm
|
pax3.L
|
|
laevis
|
NF stage 21
|
neuroectoderm
,
neural crest
,
neural plate
,
preplacodal ectoderm
|
Display additional annotations [+]
Gene |
Clone |
Species |
Stages |
Anatomy |
zic1
|
|
laevis
|
NF stage 19
|
neuroectoderm
,
neural plate
,
cranial neural crest
|
zic1
|
|
laevis
|
NF stage 14
|
neuroectoderm
,
neural crest
,
neural plate
,
presumptive rhombomere
|
zic1
|
|
laevis
|
NF stage 18
|
neuroectoderm
,
neural crest
,
neural plate
,
presumptive rhombomere
|
zic1
|
|
laevis
|
NF stage 21
|
neuroectoderm
,
neural crest
,
neural plate
,
presumptive rhombomere
|
snai2
|
|
laevis
|
NF stage 19
|
neuroectoderm
,
neural crest
,
neural plate
,
cranial neural crest
,
neural plate border
|
vgll3
|
|
laevis
|
NF stage 15
|
neuroectoderm
,
neural plate
,
presumptive rhombomere
|
vgll3
|
|
laevis
|
NF stage 18
|
neuroectoderm
,
neural plate
,
presumptive rhombomere
|
vgll3
|
|
laevis
|
NF stage 14
|
neuroectoderm
,
neural plate
,
presumptive rhombomere
|
vgll3
|
|
laevis
|
NF stage 21
|
neuroectoderm
,
neural plate
,
presumptive rhombomere
|
vgll3
|
|
laevis
|
NF stage 19
|
neuroectoderm
,
neural plate
,
presumptive rhombomere
|
|
|
Fig. 3. Vgll3 stimulates pax3 and zic1 expression. (A) Embryos were injected with
v3MO or vgll3 mRNA and analysed at stage 19 for pax3 and zic1 expression.
Defects of CNC lateral streams and trigeminal placode are shown by arrows and
arrowhead respectively. (B) ISH for vgll3 and pax3 shows overlapping expression at
the r2 level (blue arrow). (C) Stage 19 embryos injected with pax3GR/zic1GR
mRNAs show a faint vgll3 and a strong snail2 expression after dexamethasone
treatment (+dex). (D) cMO/LacZ mRNA control and quantification. Three
independent experiments were performed (n=40). (*), injected side. All views are
dorsal-anterior. A dotted line indicates the midline embryo. (E) ISH for vgll3 and zic1 shows overlapping expression at the r2 level (blue arrow). Zic1 expression in stage
21 embryo. |