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Fig. 5. XFlop is necessary for cortical actin assembly. Two antisense oligos (1s
and 5s) were used to deplete the maternal store of XFlop mRNA in
oocytes, which were then fertilized to assess the effect of maternal XFlop
depletion. (A) The efficiency of depletion of XFlop mRNA by oligos 1s
(10 ng/oocyte) and 5s (8 ng/oocyte); XFlop mRNA is not resynthesized
before the gastrula stage. Oocyte, st.9 and st.10.25 represent the levels of
XFlop mRNA, normalized to the level of ODC mRNA at the oocyte, late
blastula and early gastrula stages. 1s and 5s represent the levels at the same
stages after injection of oligo 1s and 5s, respectively. XFlop-depleted
embryos were flattened when compared with control embryos, as shown from above
(B), and from the side of bisected embryos (C). (D) XFlop depletion resulted
in slower wound healing. Quantitation of this effect is shown, and is compared
with that caused by the overexpression of XFlop in the same experiment, shown
in Fig. 3E. (E) The effect of
XFlop depletion on cortical actin at stage 9. XFlop depletion reduced the
overall amount of cortical actin (upper panel, low-magnification confocal
images; middle panel, pixel intensity; lower panel, high-magnification
confocal images). (F) The decrease of the overall amount of the cortical actin
caused by XFlop depletion is significant (P<0.001, n=5
independent experiments). (G) The effects of XFlop depletion on F-actin in
single dissociated cells further indicate that XFlop is required for the
cortical actin assembly. Twenty control and 28 XFlop-depleted cells were
imaged and the pixel intensity was measured. (H) Quantitative analysis shows
the decrease of the pixel intensity is significant (P<0.01).
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