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Fig. 1. Alterations in somite formation and expression of somitogenesisrelated
genes in Bowline-deficient embryos. (A) Western blotting analysis
of Bowline-myc protein. Two-cell stage embryos were injected in the
animal hemisphere with 200 pg of RNA encoding myc-tagged Bowline
(lanes: 1–3) or myc-tagged Bowline2 (lanes: 4–6), and 20 ng of bowline
MO (lane 2), bowline2 MO (lane 5), or 20 ng of CoMO (lanes: 3 and 6).
The 9E10 anti-myc antibody was used for immunoblotting. As a positive
control, a monoclonal antibody was used to detect a-tubulin. (B,C)
Embryos at 16-cell-stage were injected at equatorial region of 1 blastomere
(V2.2 or D2.2 position [27]) with combination of bowline MO (5 ng) and
bowline2 MO (5 ng) (BlnMO) or CoMO (10 ng) as indicated. RNA
encoding the lineage tracer b-gal was co-injected to identify the injected
side (red staining). Injected sides are indicated as ‘‘inj’’. (B) Eembryos were
fixed and stained with Red-gal and hematoxylin at stage 28–32. Embryos
are oriented with anterior to the left. Frontal sections. Bars, 100 nm. The
arrowheads indicate cells introduced with MOs. (C) Changes in expression
patterns of somitomeric markers. Embryos were analyzed for expression
of Thy1, X-Delta-2, bowline intron, X-hairy-2, or X-Delta-1 by wholemount
in situ hybridization at stage 20–21. Dorsal views are shown with
anterior towards the top. The arrowheads indicate positions where the
gene expression patterns are different from those of the uninjected side.
The arrows indicate ectopic expression of Thy1 between stripes. The
dotted lines indicate regions of Thy1 or bowline expression along the
anteroposterior axis (black line, uninjected sides; red line, injected sides).
(For interpretation of the references in color in this figure legend, the
reader is referred to the web version of this article.) |