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Fig. 6. Somite transplants restore the normal PMN subtype pattern in
ntl;spt mutants. (A) Schematic of blastula stage transplants. (B-D,F)
Islet antibody + islet2 in situ hybridization + anti-fluorescein
antibody staining at 18-22 hpf. Islet antibody staining is nuclear and brown;
islet2 staining is blue and cytoplasmic; red staining is fluorescent
and shows wild-type donor cells that contain fluorescein dextran. Brown-only
cells (*) express only islet1 and hence are MiPs; blue +
brown cells express islet2 and possibly also islet1, and are
therefore CaPs or hybrid PMNs. (B) ntl;spt MO-injected host embryo
with its spinal cord completely filled with wild-type donor cells but devoid
of wild-type somite cells. No MiPs (brown-only cells) are present in this
embryo, so the PMNs probably still all have a hybrid identity. (C)
Bright-field microscopy and (D) fluorescence microscopy of the same
cross-section of another ntl;spt MO-injected host embryo with its
spinal cord completely filled with wild-type donor cells but devoid of
wild-type somite cells. Two triple-labeled PMNs are indicated with arrowheads.
(E) Schematic of whole somite transplants. (F) ntl;spt MO-injected
host embryo with transplanted wild-type somites. Several MiPs (brown-only
cells; *) are present adjacent to the wild-type somites (red).
These MiPs are separated by blue + brown cells that are probably CaPs. Insert
shows a different ntl;spt host embryo with transplanted wild-type
somites; in this case, the somite boundaries are clearly visible (broken
lines). As in wild-type embryos, MiPs were adjacent to these somite
boundaries. Scale bar: 50 µm.
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