Jerry R , Sullivan-Brown J , Yoder MD .

	Fig. 1. Gtpbp10 is a member of the Obg family of G-proteins and is conserved among metazoans. A) Amino acid alignment of Xenopus laevis Gtpbp10 (Xl) with the prokaryotic ObgE protein from Bacillus subtilis (Bs). Glycine residues are conserved in the N-terminal glycine-rich domain (ObgE fold), amino acids 1–158 (red bar). Five conserved ‘G-box’ domains are denoted by black lines as G1-G5. Consensus sequences are as follows: G-box 1: GXXXXGK [T/S]; G-box 2: conserved T residue at position 177 (197 in B. subtilis); G-box 3: DXXG; G-box 4: [N/T]KXD; and G-box 5: [C/S]A [K/L/T]. The conserved Switch I and Switch II nucleotide binding and hydrolysis domains are denoted by red and green boxes, respectively. B) Phylogenetic tree representing maximum likelihood analysis of the P-loop GTPases Gtpbp10/ObgE (blue), Drg1/ObgE (yellow), Srp54/Ffh (purple), and EF-1a/EF-TU (orange). The outgroup is represented by B. subtilis FtsZ (Bs FtsZ, green). Species abbreviations: Ce: Caenorhabitis elegans, Ci: Ciona intestinalis, Ct: Capitella teleta, Dm: Drosophila melanogaster, Dr: Danio rerio, Ob_EF1a, Hs: Homo sapien, Hv: Hydra vulgaris, Mm: Mus musculus, Ob: Octopus bimaculoides, Xt: Xenopus tropicalis. Tree was assembled using PhyML 3.0 - ATGC-Montpellier (bootstrapping was set at 100). All alignments were created using Clustal Omega (EMBL-EBI). C) Amino acid alignment and comparison of Gtpbp10 from 6 metazoans. The Switch I and Switch II domains highlighted as above. D) Percent similarity and identity of selected Gtpbp10 orthologs from B (compared to Xenopus laevis), ObgE from B. subtilis shown for comparison.
	Fig. 2. GTPBP10 mRNA is expressed throughout development. RT-PCR was performed using RNA isolated from the indicated developmental stages. gtpbp10 migrates at 248bp, while ODC migrates at 221bp (200bp band indicated with an asterisk). ODC serves as a positive loading control. ‘-RT’ represents cDNA synthesis in the absence of reverse transcriptase (negative control).
	Fig. 3. Whole-mount in situ hybridization (WISH) indicates no early expression of gtpbp10. gtpbp10 antisense riboprobe was used for in situ hybridization on embryos of the indicated stages. A and B) Animal and vegetal views of stage 9 embryos (respectively), animal cap was removed for imaging in A. C and D) Dorsal-Vegetal view of stage 11 and 12 embryos, respectively, dorsal side is pointing up. E) Dorsal view of stage 18 embryo along the anterior-posterior axis (anterior up), the pigmented neural fold is shown, with incomplete closure in the anterior neuroectoderm.
	Fig. 4. The expression of gtpbp10 in late stages of development shown by whole-mount in situ hybridization (WISH). gtpbp10 antisense riboprobe was used for in situ hybridization on A) Early tail bud stage embryo (stage 28) and B) Late tail-bud stage (stage 36). A′ and B′ are transverse sections of the embryos shown in A and B at the level of the dotted white lines. Structures labeled as follows: notochord (n), neural tube (nt), forebrain (fb), hindbrain (hb), pronephros (pn), branchial arches (ba). * denotes the glomerulus and proximal tubules, while ‘t’ indicates the distal tubules. C) gtpbp10 sense probe shows background staining in the anterior branchial arches, otic vesicle, eye, and fore-, mid-, and hindbrain. Staining was also observed in somitic tissue (so) along the anterior-posterior axis.
	gtpbp10 (GTP-binding protein 10 (putative)) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 36, lateral view, anterior left, dorsal up.