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Figure S5. Temperature and MO Dosage Effects on Brachyury Phenotype and
Immune Response Related Gene Transcription, Related to Figure 7
(A) WMISH for tp53 of late tailbud embryos injected with 4.5 or 18 ng of the t/t2 MO
mix and developed at 22 ºC or 28.5ºC. *, Remark: Increasing incubation temperature
also slightly up-regulated tp53 in the absence of any MO (confirmed by RT-qPCR,
data not shown). (B) Multi-probe WMISH for various mesoderm cell lineage and
derivative markers (cav1, notochord; hoxd8, pronephros; myh6, heart; tal1, ventral
blood island; tbx6, paraxial mesoderm) and single WMISH for tp53, tp53inp1 and
c3ar1 of mid-tailbud (stage 26) and late tailbud embryos (stage 34) injected with 6 or
18 ng of the t/t2 MO mix. White arrowheads point to the expression domains of tbx6
and cav1 that were not maintained in embryos without functional Brachyury. (C)
WMISH for tp53inp1 on wild-type (or t/t2 heterozygous) and t/t2 null mutant embryos
as well as embryos injected with 1 and 3 ng MO. The embryos were developed to
late tailbud stage 34 at 25-26ºC. Scale bar, 0.5 mm. |
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Figure S3. Increased Transcription of tp53 Depending on GC Content of MO
Does Not Cause More Apoptosis, Related to Figure 3 and 4
(A) TUNEL assay on morphants and sibling embryos from double heterozygous t
-/+t2+/-
parents. DNase-treated wild-type embryos were used as positive controls. (B) Single
WMISH for tp53 and multi-probe WMISH for various mesoderm cell lineage and
derivative markers (cav1, notochord; hoxd8, pronephros; myh6, heart; tal1, ventral
blood island; tbx6, paraxial mesoderm) of late tailbud embryos injected with single
MOs or tracer sulforhodamine-dextran. Scale bar, 0.5 mm |
