ZFIN ID: ZDB-FIG-070619-36
Qian et al., 2007 - Distinct Functions for Different scl Isoforms in Zebrafish Primitive and Definitive Hematopoiesis. PLoS Biology   5(5):e132 Full text @ PLoS Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 14-19 somites to Prim-15

Fig. 7 Differences in Protein Expression Levels of Scl-α and -β Confer Their Distinct Functions (A–H) βe1-globin and c-myb expression in scl-sp morphants is rescued by scl-α and -β mRNA. (A–D) show WISH of βe1-globin in 20-hpf control embryos (A), scl-sp morphants (B), scl-sp morphants injected with scl-α mRNA (C), and scl-sp morphants injected with scl-β mRNA (D). (E–H) show WISH of c-myb in 30-hpf control embryos (E), scl-sp morphants (F), scl-sp morphants injected with scl-α mRNA (G), and scl-sp morphants injected with scl-β mRNA (H). Insets in (E–H) are high-magnification (20×) views of the ventral wall of DA, indicated by black arrowhead. In all panels, embryos are in lateral view with anterior to the left. (I) Immunoblotting with Ab-Scl-C antiserum shows the expression levels of Scl-α and Scl-β proteins in wild-type (wt) embryos, clo mutant embryos, and scl-α morphants. Although Scl-β protein was not detectable, immunoblotting of whole embryo protein extracts (50 μg for each sample) showed that Scl-α protein expression increased as embryos developed from the 18-somite (lane 1) to 22-somite (lane 4) stage. The clo mutant embryos (lane 3) and scl-α morphants (lane 2), in which Scl-α protein was not detected, were used as the controls to distinguish the Scl-α protein (arrow) and nonspecific band (asterisk). (J) Immunoblotting with Ab-Scl-C antiserum shows the expression levels of Scl-α and Scl-β proteins in transfected COS7 cells. Western blotting of whole cell protein extracts (10 μg for each sample) prepared from COS7 cells transfected with full-length scl-α, scl-β, or blank vector constructs revealed that the protein level of Scl-α was much higher than that of Scl-β. RT-PCR analysis (bottom) showed similar RNA levels of scl-α and -β in these transfected cells. Tubulin and GAPDH were used as controls for Western blot and PCR, respectively. (K) Immunoblotting with Ab-Scl-C antiserum shows the expression levels of Scl-α and Scl-β proteins in the embryos injected with in vitro synthesized scl-α or -β mRNA. Western blotting of whole embryo protein extracts (50 μg for each sample) from embryos injected with 500 pg of scl-α or -β mRNA showed that, at 3 h post-injection, the Scl-β protein was detected as being at a comparable level to that of Scl-α but dramatically reduced by 4 h post-injection.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
hbbe1.1 WT control Prim-15 ventral wall of dorsal aorta ISH
WT standard conditions Prim-15 posterior lateral mesoderm ISH
myb WT control Prim-15 intermediate cell mass of mesoderm ISH
WT standard conditions Prim-15 posterior lateral mesoderm ISH
tal1 WT control 14-19 somites whole organism IHC
20-25 somites whole organism IHC
WT + MO7-tal1 standard conditions 20-25 somites Not Detected whole organism IHC
npas4ls5/s5 standard conditions 20-25 somites Not Detected whole organism IHC
Antibody Labeling Details No data available
Phenotype Details No data available
Acknowledgments:
ZFIN wishes to thank the journal PLoS Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Biol.