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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. |
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Gene |
Clone |
Species |
Stages |
Anatomy |
tubb2b
|
|
laevis
|
NF stage 19
|
neuroectoderm
,
neural plate
,
preplacodal ectoderm
|
tubb2b
|
|
laevis
|
NF stage 20
|
neuroectoderm
,
neural plate
,
preplacodal ectoderm
|
tubb2b
|
|
laevis
|
NF stage 28
|
brain
,
central nervous system
,
trigeminal placode
,
trigeminal ganglion
,
nerve
|
neurod1
|
|
laevis
|
NF stage 18
|
neuroectoderm
,
preplacodal ectoderm
|
neurod1
|
|
laevis
|
NF stage 19
|
neuroectoderm
,
preplacodal ectoderm
|
vgll3
|
|
laevis
|
NF stage 19
|
neuroectoderm
,
neural plate
,
presumptive rhombomere
|
|
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Fig. 2. Vgll3 knockdown or overexpression impair trigeminal placode and
nerve formation. (A) Embryos injected with v3MO or v3MOsplicing (v3LMOsp and
v3SMOsp, 20 ng each) exhibit a reduced expression of islet1, neuroD and N-tubulin
in the trigeminal placodes (arrowhead). (B) Overexpression of increasing amounts of
vgll3 mRNA reduces islet1 and N-tubulin expression in stage 19 embryos. Double
ISH shows no prominent overlapping staining between vgll3 (red) and N-tubulin
(blue) (arrow). Injected side (*) was traced by lacZ staining. Gene expression were
assayed by ISH. Arrowhead indicates the trigeminal placodes. (C) Quantification of
results. Images on the top of bars indicated v3LMOsp defects rescued with increasing amounts of vgll3 mRNA injections. Three independent experiments were
performed. The number of embryos analysed is indicated on the top of each bar.
Views are dorsal-anterior excepted for lateral views for stage 28 embryos. |