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Figure 1. Temporal and spatial expression pattern of the Hoxc6 isoforms. A: Structure of Hoxc6 isoforms in Xenopus laevis. Hoxc6 isoforms derive from 2 different promoters. The distal promoter (PR I) leads to a 2.2-Kb precursor transcript, coding for the short form (SF) protein of 152 aa. The proximal promoter encodes a 1.8-Kb transcript leading to the long form (LF) protein of 234 aa. Black boxes indicate the open reading frame, grey boxes are the homeodomain. Red bars indicate the binding sites of the morpholino-oligonucleotides. The blue and green arrows show the primer sites that served for the RT-PCR and to generate the probes for in situ hybridization. B: Temporal expression pattern of Hoxc6 isoforms. RT-PCR was carried out using embryos at the indicated stages from 1 to 27. H4 is used as a loading control; -RT is shown for stage 12 RNA. C-K: SF expression patterns from beginning of gastrulation until tadpole stages. Stages are indicated by the numbers. L-T: LF pattern from beginning of gastrulation until tadpole stages. The arrowheads in K and T indicate the location of the pronephric anlage. The squared brackets indicate the different distances between the anteriormost border of expression of each isoform and the otic vesicle area. |
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Figure 2. Tissue localisation of Hoxc6 isoforms in cross-sections. A: SF expression in the somitic tissue and neural tube. B: Expression of LF in somitic tissue and neural tube; in addition, LF is expressed in the pronephric anlage (arrowheads). C, D: Close-ups of the neural tubes shown in A and B. Expression of the Hoxc6 isoforms is detected in all cells of the neural tube with exception of the floorplate. E: MyoD expression in the somites. F: Xlim-1 expression in the neural tube and in the pronephric anlage (arrowheads). G: FoxF1 expression in the lateral plate mesodem. H: Position of the shown sections within stage-27 embryos, and localization of different tissues in the sections. Neural tube (blue); somite mesoderm (red); notochord (brown); endoderm (yellow); lateral plate mesoderm (dark orange); the pronephric anlage (light orange). I: Cross-section at the level of the branchial arches (br) as indicated in the inlay. Expression of SF is found in the neural tube (ne), even though it was not visible in whole mount in situ hybridization. J: Expression of LF in the neural tube at the same level. |
