FIGURE

Fig. S3

ID

ZDB-FIG-081104-1

Publication

Leung et al., 2008 - The orphan G protein-coupled receptor 161 is required for left-right patterning

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Fig. S3

Translational blocking by gpr161-MO morpholino detected by in vitro cell lysate assay. (A) Two translational blocking morpholinos, targeted against the 5′UTR upstream of the ATG start codon of zebrafish gpr161 open-reading frame, which was fused to V5-6xHistidine epitope. (B,C) Dose dependent inhibition of Gpr161 translation by two independent gpr161 morpholinos (gpr161-MO#36 and gpr161-MO#24) using in vitro cell lysate. No inhibition by 5-base mismatch control morpholino (mismatch-MO#28). Anti-V5 antibody was used to detect the in vitro translation (B) of Gpr161-V5 fusion protein and band intensity (C) was quantitated as % translation compared to control in the graph. No morpholino as control (100%), 10 μM MO#24 (7%), 1 μM MO#24 (56%), 0.1 μM MO#24 (84%), 10 μM MO#28 (79%), 1 μM MO#28 (92%), 0.1 μM MO#28 (79%), 10 μM MO#36 (0%), 1 μM MO#36 (13%) and 0.1 μM MO#36 (40%). (D) Both gpr161 morpholinos (MO#24 and MO#36) gave similar cardiac looping defect. All scale bars were 100 μm.

Expression Data

Gene:	myl7
Fish:	WT WT + MO1-gpr161b WT + MO2-gpr161b
Knockdown Reagents:	MO1-gpr161b MO2-gpr161b
Anatomical Terms:	presumptive atrium heart tube presumptive cardiac ventricle heart tube
Stage:	Prim-25

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	WT + MO1-gpr161b WT + MO2-gpr161b
Knockdown Reagents:	MO1-gpr161b MO2-gpr161b
Observed In:	atrium cardiac ventricle heart looping
Stage:	Prim-25

Phenotype Detail

Acknowledgments

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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.