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Fig. 1. SoxB1 protein encoded from mRNA microinjected at the one-cell stage does
not persist in vegetal cells. (A) Translation of SoxB1 mRNA is efficiently
blocked in embryos that develop from zygotes injected with SoxB1MO, as shown
by the absence of staining with an antibody specific for SoxB1 (green, compare
with B and C). Red staining detects the 6e10 extracellular epitope produced by
primary mesenchyme cells. This embryo is at the temporal equivalent of prism
stage but has failed to differentiate an archenteron
(Kenny et al., 2003). (B)
Co-injection of the SoxB1 morpholino and MO-immune SoxB1 synthetic
mRNA rescues differentiation of gut and coelomic rudiments (white arrowheads).
SoxB1 protein translated from the microinjected mRNA (green) accumulates and
persists in the ectoderm, but does not persist in secondary mesenchyme (white
arrowheads) or endoderm (except for low levels in the foregut). (C) An embryo
at the hatched blastula stage treated as in B also clears SoxB1 protein (green
signal, left) from vegetal blastomeres, despite the fact that the
microinjected mRNA is present in them, as shown by whole-mount in situ
hybridization to SoxB1 mRNA in the same embryo (blue signal, right).
The embryo is stained for SoxB1 (FITC) and DNA (DAPI); the merged fluorescent
signals over SoxB1-positive nuclei are blue-green, whereas those of
SoxB1-negative nuclei are blue. Immunofluorescence images are shown on the
left; DIC images, right. A and V indicate animal and vegetal poles,
respectively. Scale bars: in B, 20 µm for A,B; in C, 20 µm.
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