ZFIN ID: ZDB-FIG-130524-12
Chen et al., 2013 - Snx3 Regulates Recycling of the Transferrin Receptor and Iron Assimilation. Cell Metabolism   17(3):343-352 Full text @ Cell Metab.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: 2-cell to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Protruding-mouth

Fig. S2

Morpholino-mediated knockdown of snx3 in zebrafish embryos specifically impairs heme production and erythroid lineage differentiation. (A) The expression profile of snx3 in different stages of early zebrafish embryos. snx3 zygotic transcripts are enriched in the ventral blood islands (intermediate cell mass, ICM) from around 10-somite stage, a pattern similar to that of tfr1a. In contrast to tfr1a, snx3 is expressed as a maternal transcript between 2- and 1000-cell stages. The erythroid transcription factor gata1 is presented as a control for primitive hematopoiesis. (B) MO1 and MO2 were designed to target the translational initiation site and the second exonintron boundary in snx3, respectively. (C) RT-PCR using primer set shown in panel (B) confirmed the presence of altered snx3 mRNA spliceforms in zebrafish embryos injected with MO2. (D) Whole-mount in situ hybridization indicated that initial specification of erythropoiesis (gata1) and myelopoiesis (myeloperoxidase or mpo) was not affected in the snx3 morphants. (E) Whole-mount in situ hybridization in snx3-morphant embryos showed normal and reduced expression of βe1-globin at 48 hpf and 72 hpf, respectively.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
gata1a AB control Prim-5 myeloid lineage restricted progenitor cell ISH
Protruding-mouth myeloid lineage restricted progenitor cell ISH
AB standard conditions 2-cell Not Detected whole organism ISH
1k-cell Not Detected whole organism ISH
10-13 somites ventral lateral mesoderm ISH
14-19 somites intermediate cell mass of mesoderm ISH
Prim-5 blood island ISH
AB + MO1-snx3 standard conditions Prim-5 myeloid lineage restricted progenitor cell ISH
Protruding-mouth myeloid lineage restricted progenitor cell ISH
AB + MO2-snx3 standard conditions Prim-5 myeloid lineage restricted progenitor cell ISH
Protruding-mouth myeloid lineage restricted progenitor cell ISH
hbbe1.1 AB control Long-pec nucleate erythrocyte ISH
Protruding-mouth nucleate erythrocyte ISH
AB + MO1-snx3 standard conditions Long-pec nucleate erythrocyte ISH
Protruding-mouth nucleate erythrocyte ISH
AB + MO2-snx3 standard conditions Long-pec nucleate erythrocyte ISH
Protruding-mouth nucleate erythrocyte ISH
mpx AB control Prim-5 erythroid progenitor cell ISH
Protruding-mouth erythroid progenitor cell ISH
AB + MO1-snx3 standard conditions Prim-5 erythroid progenitor cell ISH
Protruding-mouth erythroid progenitor cell ISH
AB + MO2-snx3 standard conditions Prim-5 erythroid progenitor cell ISH
Protruding-mouth erythroid progenitor cell ISH
snx3 AB standard conditions 2-cell whole organism ISH
1k-cell whole organism ISH
10-13 somites whole organism ISH
14-19 somites intermediate cell mass of mesoderm ISH
Prim-5 blood island ISH
tfr1a AB standard conditions 2-cell whole organism ISH
1k-cell whole organism ISH
10-13 somites whole organism ISH
14-19 somites intermediate cell mass of mesoderm ISH
Prim-5 blood island ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO1-snx3 standard conditions Protruding-mouth nucleate erythrocyte decreased amount, abnormal
AB + MO2-snx3 standard conditions Protruding-mouth nucleate erythrocyte decreased amount, abnormal
Acknowledgments:
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Reprinted from Cell Metabolism, 17(3), Chen, C., Garcia-Santos, D., Ishikawa, Y., Seguin, A., Li, L., Fegan, K.H., Hildick-Smith, G.J., Shah, D.I., Cooney, J.D., Chen, W., King, M.J., Yien, Y.Y., Schultz, I.J., Anderson, H., Dalton, A.J., Freedman, M.L., Kingsley, P.D., Palis, J., Hattangadi, S.M., Lodish, H.F., Ward, D.M., Kaplan, J., Maeda, T., Ponka, P., and Paw, B.H., Snx3 Regulates Recycling of the Transferrin Receptor and Iron Assimilation, 343-352, Copyright (2013) with permission from Elsevier. Full text @ Cell Metab.