|
|
|
|
|
|
|
||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | ||||
| ||||||||||||||||||||
Files in this Data Supplement:
Fig S1. cdx4, but not cdx1a, is expressed in the prospective spinal cord territory before and during somitogenesis. Expression of cdx1a (A,A′,C,E) and cdx4 (B,B′,D,F) (purple) at the tailbud (A,A′,B,B′), 6-somite (C,D) and 10-somite (E,F) stages of development. Arrow indicates anterior limit of cdx4 expression, as compared with adjacent adaxial cells and somites (s) labeled for myod (red). Flat-mounted embryos, anterior to the left, except A′ and B′ which are lateral views of whole embryos, anterior to the top. Scale bars:100 μm. (G) Change in the anterior limit of cdx4 expression in the CNS as a function of time. Embryos at different developmental stages (hpf, hours post-fertilization, x-axis) were hybridized for cdx4 and myod and the anterior limit of cdx4 expression in the CNS analyzed with respect to adjacent somites (y-axis). Each point is the average of at least ten embryos; s.e. indicated with bars.
Fig. S2. Retinoic acid pathway is not affected in cdx1a/cdx4-deficient embryos at the end of gastrulation. Expression of genes encoding the retinoic acid (RA) -synthesizing enzyme Raldh2 (also known as Aldh1a2 − ZFIN) (A,D), Retinoic acid receptor α (RARα; also known as Raraa − ZFIN) (B,E) and the RA-degrading enzyme Cyp26a1 (C,F) in wild-type (A-C) and cdx1a/cdx4-deficient embryos (D-F) at the end of gastrulation (tailbud stage, 10 hpf). Expression of these genes in the paraxial mesoderm, head and tail regions of cdx1a/cdx4-deficient embryos is similar to that seen in wild-type siblings. More than 40 embryos from two independent experiments were analyzed, all displaying the expression patterns shown. Embryos are shown in dorsal (A,D) or lateral (B,C,E,F) views, anterior to the top. Scale bar: 100 μm.
Fig. S3. Heat-induced rapid and generalized expression of phsp70:cdx4 transgene. Expression of cdx4 (purple) and myod (red in adaxial cells) in wild-type embryos (A) and their siblings carrying one copy of a phsp70:cdx4 transgene (B) that were heat shocked for 1 hour at 37°C starting at the three-somite stage (11 hpf) and immediately fixed. From three crosses between a wild-type and a transgenic heterozygous fish, 48% of embryos showed ubiquitous cdx4 expression (n=428, B). Upon genotyping, 100% of embryos expressing cdx4 ubiquitously were also positive for the transgene and 100% of embryos with a normal cdx4 pattern of expression tested negative for the transgene (n=20 for each class). Embryos are shown in dorsal view, anterior to the left. Scale bars: 100 μm.
Fig. S4. Rescue of blood marker gata1 expression in cdx1a/cdx4-deficient embryos by hoxa9a over-expression. 20-somite stage (19 hpf) wild-type (A), cdx1a/cdx4-deficient (B) and hoxa9a-injected cdx1a/cdx4-deficient (C, 25 pg mRNA) embryos stained for the blood marker gata1 (purple). A minimum of 15 injected embryos in three independent experiments were analyzed, with more than 90% of embryos displaying the phenotypes shown. Scale bar: 100 μm.
| ||||||||||||||||||||
