FIGURE

Fig. S3

ID

ZDB-FIG-190624-2

Publication

van der Klaauw et al., 2019 - Human Semaphorin 3 Variants Link Melanocortin Circuit Development and Energy Balance

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Fig. S3

Generation and Characterization of Semaphorin-Neuropilin-Plexin Deletion Mutants in Zebrafish, Related to Figure 3

(A) Schematic illustrating the mutagenesis strategy to target sema3, plxna and nrpgenes in zebrafish. Two-five sgRNAs were generated to mutagenize each zebrafish gene. Only sgRNAs verified to induce mutagenesis were injected into one-cell stage zebrafish embryos. Zebrafish were raised to ∼30 days post fertilization and fish length (mm), weight (mg) and % body fat were quantified.

(B) Results on fish length (mm), weight (mg) and % body fat for all deletion mutants (summarized in Figure 3B).

(C) Microphotographs and quantification of the density of α-melanocyte-stimulating hormone (αMSH) (red) and agouti-related peptide (AgRP) (green) immunoreactive (IR) fibers innervating the preoptic area (POA), anterior tuberal nucleus of hypothalamus (ATN), and lateral hypothalamic nucleus (LH) of 35-day-old wild-type zebrafish overexpressing NRP1 and NRP2; ^∗p < 0.05 in one-sample t tests.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	WIK + CRISPR1-nrp2b + CRISPR2-nrp2b WIK + CRISPR1-plxna2 + CRISPR2-plxna2 WIK + CRISPR1-plxna3 + CRISPR2-plxna3 WIK + CRISPR1-plxna4 + CRISPR2-plxna4 WIK + CRISPR1-sema3aa + CRISPR2-sema3aa WIK + CRISPR1-sema3ab + CRISPR2-sema3ab WIK + CRISPR1-sema3fa + CRISPR2-sema3fa WIK + CRISPR2-sema3d + CRISPR3-sema3d
Knockdown Reagents:	CRISPR1-nrp2b CRISPR1-plxna2 CRISPR1-plxna3 CRISPR1-plxna4 CRISPR1-sema3aa CRISPR1-sema3ab CRISPR1-sema3fa CRISPR2-nrp2b CRISPR2-plxna2 CRISPR2-plxna3 CRISPR2-plxna4 CRISPR2-sema3aa CRISPR2-sema3ab CRISPR2-sema3d CRISPR2-sema3fa CRISPR3-sema3d
Observed In:	adipose tissue whole organism
Stage:	Days 30-44

Phenotype Detail

Acknowledgments

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Reprinted from Cell, 176(4), van der Klaauw, A.A., Croizier, S., Mendes de Oliveira, E., Stadler, L.K.J., Park, S., Kong, Y., Banton, M.C., Tandon, P., Hendricks, A.E., Keogh, J.M., Riley, S.E., Papadia, S., Henning, E., Bounds, R., Bochukova, E.G., Mistry, V., O'Rahilly, S., Simerly, R.B., INTERVAL, UK10K Consortium, Minchin, J.E.N., Barroso, I., Jones, E.Y., Bouret, S.G., Farooqi, I.S., Human Semaphorin 3 Variants Link Melanocortin Circuit Development and Energy Balance, 729-742.e18, Copyright (2019) with permission from Elsevier. Full text @ Cell