RNA-FISH analyses of Xist in cumulus cell-derived cloned embryos
(A and B), female IVF embryos (C), and female ES cell-derived embryos (D). The right
panels show the ratios of blastomeres classified according to the number of cloud or
pinpoint signals of Xist RNA. A: At the morula stage (72 h after
activation), the Xist siRNA treatment significantly increased the
proportion of blastomeres without cloud signals (counting those with two pinpoints +
one pinpoint + no signal) (19 vs. 76% on average; P<0.005). B:
At the blastocyst stage (96 h after activation), there was no significant difference
in the proportion of blastomeres without cloud signals between the control and
Xist-specific siRNA groups (40 vs. 41%;
P>0.05). C: In normally fertilized female embryos, the majority of blastomeres
showed expression of one cloud signal in controls and one pinpoint signal in
Xist-specific siRNA-injected embryos. D: In embryos cloned from
female ES cells, the expression patterns or Xist with or without
Xist-specific siRNA treatment were similar to those found in
cumulus cell-derived cloned embryos (see A). (Scale bar, 50 μm).