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Fig. 1. TC/TC homozygous mutants produce a single
but persistent mRNA transcript and transient protein. (A) RT-PCR
reactions for +/+ (LHF to 8-s), TC/+ (LHF to 8-s) and
TC/TC (LHF, 5-s) conceptuses. A single
T transcript was detected in +/+ samples, a strong T and a
faint TC band were detected at all stages in
TC/+ samples, and a single TC
transcript was detected in TC/TC samples. Minus
reverse-transcriptase (-RT) control was negative for all reactions. M
indicates marker, a 100 bp gene ruler. ß-actin RT-PCR product was used as
a loading control for each corresponding T RT-PCR reaction. (B)
Immunohistochemical detection of T and TC (brown) in +/+ (a,b) and
TC/TC (c,d) conceptuses in sagittally oriented
histological sections. T (a) and TC (c) were localized to the
primitive streak (ps) at the EB stage. At LHF, T (b) was localized to the
allantoic core and primitive streak, and, with the exception of the
ectoplacental endoderm (not shown), TC (d) was not detected in the
TC/TC conceptus. The slanted black line in b
indicates the previously defined boundary between the allantois and posterior
primitive streak (Downs and Harmann,
1997). (e-g) Transverse immunohistochemical sections (6 µm)
through a 2-s allantois to highlight the T-defined core domain. The total
length of this allantois was 258 µm after fixation. (e) The distalmost T(+)
section at 102 µm (39.5% of the total fixed allantoic length). (f) A
mid-region section at 48 µm. (g) The most proximal section at 6 µm. ac,
amniotic cavity; al, allantois; am, amnion; bi, yolk sac blood island; eve,
embryonic visceral endoderm; ps, primitive streak; x, exocoelomic cavity; xve,
extraembryonic visceral endoderm. Scale bars: in d, 100 µm for a-d; in g,
100 µm for e-g.