| |
Figure 5.
RI in wnt11b Is Responsible for Somite Defects in tra2b Morphants
(A) Diagram showing X. laevis wnt11b gene structure and the wnt11b-in4 MO (red).
(B) RT-PCR on stage 19 single embryos injected with wnt11b-in4 MO or tra2b MO shows efficient retention of intron 4. The top panel shows RT-PCR with primers in wnt11b exon 4 and intron 4, and the bottom panel shows the internal control odc.
(C) ISH for mesodermal gene expression in wnt11b-in4 MO- and tra2b MO-injected embryos. Black arrows in pcdh8-stained embryos indicate the presence of somitic stripes in control and wnt11b-in4 MO-injected embryos. Red arrows in hey1-stained embryos indicate mature somites in control embryos, which are absent in wnt11b-in4 and tra2b morphants. The asterisk (∗) in hey1 samples indicates nonsomitic midline hey1 expression, which is exposed because of a neural tube closure defect. All pictures show dorsal views with anterior up.
(D and E) Quantification of ISH results, showing the mean and SD of the number of hey1+ somites (D) and pcdh8+ stripes (E); ∗∗∗ indicates that difference is statistically significant from control at p < 2.2 × 10−15 (t test). Number of embryos used for quantification: 41 (ctrl, hey1), 30 (tra2bMO, hey1), 43 (wnt11bMO, hey1), 42 (ctrl, pcdh8), 32 (tra2bMO, pcdh8), and 43 (wnt11bMO, pcdh8). Data shown are from X. laevis. |
