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supplementary figure S10. Double WISH of RarB2 and Ripply2 |
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Fig. 1. Expression of X. laevis Rarβ1 and Rarβ2.
(A-C) Whole-mount in situ hybridization of Rarβ1 and Rarβ2
mRNA expression at stage 26 in lateral (A,C) and anterior (B)
views. Rarβ2 is expressed in the hatching gland and mature
somites, with weaker expression in the eye and branchial
arches. Rarβ1 is undetectable by whole-mount in situ
hybridization. (D) QPCR showing Rarβ1 and Rarβ2 gene
expression averaging two biological replicates over
developmental time. Error bars indicate s.e.m. The y-axis
represents 2−δCt values (adjusted for primer efficiency),
normalized to a reference gene Histone H4. |
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Fig. 2. Rarβ2 is induced by TTNPB and is regulated by RARα and/or
RARγ. (A) QPCR showing Rarα1, Rarα2, Rarβ2, Rarγ1 and Rarγ2 expression
in embryos treated at stage 7/8 with 1 μM TTNPB, 1 μM AGN193109 or vehicle
(0.1% ethanol) and collected at tailbud stage. The y-axis represents 2−δδCt
values normalized to Eef1a1 and expressed as fold induction relative to control
vehicle (n=5 biological replicates) using standard propagation of error
(Bevington and Robinson, 2003). Error bars indicate s.e.m. An unpaired t-test
in GraphPad Prism v5.0 is reported (*P≤0.05, **P≤0.01, ***P≤0.001).
(B) Embryos were injected unilaterally at the 2- or 4-cell stage with 6.6 ng Rarα
MOs or Rarγ MOs. The injected side is indicated by magenta β-gal lineage
tracer. Rarα MOs and Rarγ MOs knock down the expression of Rarβ2 (α MOs,
13/13 embryos; γ MOs, 8/8) at tailbud stage. Embryos are shown in dorsal view
with anterior on the left. Midline is indicated by a broken green line. |
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supplementary figure 4 |
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rarb (retinoic acid receptor beta) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up. |
