|
Fig. S2 Phylogram and RT-PCR analysis of the zebrafish gpr161. (A) Phylogram analysis revealed zebrafish Gpr161 protein is highly similar to the human counterpart within the purine receptor cluster branch of the δ-group of RHODOPSIN receptors. The protein sequences were aligned using T-coffee (http://igs-server.cnrs-mrs.fr/Tcoffee/tcoffee_cgi/index.cgi) and the phylogenetic tree was constructed using the Treetop program with a bootstrap value of 100and the phylogenetic tree was constructed using the Treetop program with a bootstrap value of 100 (http://www.genebee.msu.su/services/phtree_reduced.html). Genbank accession numbers for human C3AR (NP_004045), human GPR103 (NP_937822), humanTRHR (NP_003292), human P2RY11 (NM_002566), human GPR82 (NP_543007), human GPR18 (CAI14306), human EBI2 (P32249), human PTAFR (NP_000943), human GPR101 (NP_473362), human GPR161 (CAI22623), human RGR (AAI01111) and zebrafish Gpr161 (EU090912).(B) RT-PCR detection of the maternal zebrafish gpr161 transcript at 1 cell stage and zygotic transcript at sphere stage (4 hpf) onward to 96 hpf, for 26 and 30 PCR cycles. actin RT-PCR was used as RNA loading control, for 22 PCR cycles. (C) gpr161 expression in 1 dpf to 3 dpf zebrafish embryos. (D) Negative control of using sense probe of gpr161. All scale bars were 100 μm.
Reprinted from Developmental Biology, 323(1), Leung, T., Humbert, J.E., Stauffer, A.M., Giger, K.E., Chen, H., Tsai, H.J., Wang, C., Mirshahi, T., and Robishaw, J.D., The orphan G protein-coupled receptor 161 is required for left-right patterning, 31-40, Copyright (2008) with permission from Elsevier. Full text @ Dev. Biol.