ZFIN ID: ZDB-FIG-200506-7
Guo et al., 2020 - Function of Arl4aa in the Initiation of Hematopoiesis in Zebrafish by Maintaining Golgi Complex Integrity in Hemogenic Endothelium. Stem Cell Reports   14(4):575-589 Full text @ Stem Cell Reports
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-15 to High-pec
PHENOTYPE:
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage Range: Prim-15 to High-pec

Fig. 6 Knockdown of arl4aa Blocked Cleavage of Notch Receptor Precursor and Downregulated Notch Signaling Pathway along DA Floor (A) Expression of Notch signaling genes in control and arl4aa morphant groups at 42 hpf (notch1, notch3, hey2, and her5) and 33hpf (gata2a and gata2b). Data are representative of three independent experiments with error bars representing the mean ± SEM. (B) Western blot showing Notch1 and NICD protein expression in control, arl4aa morphant embryos, and DAPT-treated embryos at 42 hpf. The numbers indicate the image intensities of immunoblotting relative to those of tubulin in each embryo group. The intensity in the controls was set arbitrarily as 1. The results are representative of at least three experiments. DAPT, N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester. (C) Confocal microscope imaging of double transgenic tp1:eGFP; kdrl:mCherry embryos injected with arl4aa MO and examined at 30 hpf. Arrowheads indicate cells in the floor of the DA with active Notch signaling. Scale bar, 100 μm. (D) Enumeration of tp1 + kdrl + HSCs in the floor of DA (n = 8 embryos per group, p < 0.001). Data are representative of three independent experiments with error bars representing the mean ± SEM. (E) hsp70l:Gal4; UAS:NICD1a embryos injected with arl4aa MO were heat shocked at 14–15 somites and c-myb expression was performed at 36 hpf. Sibling embryos without NICD1a were similarly heat shocked as control. Scale bar, 250 μm. The number of embryos with typical c-myb expression as shown over the total number of embryos examined is shown in each panel.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP ci5Tg; um14Tg standard conditions Prim-15 trunk vasculature IFL
ci5Tg; um14Tg + MO1-arl4aa standard conditions Prim-15 trunk vasculature IFL
gata2a WT standard conditions Prim-15 whole organism RTPCR
WT + MO1-arl4aa standard conditions Prim-15 whole organism RTPCR
gata2b WT standard conditions Prim-15 whole organism RTPCR
WT + MO1-arl4aa standard conditions Prim-15 whole organism RTPCR
her5 WT standard conditions High-pec whole organism RTPCR
WT + MO1-arl4aa standard conditions High-pec whole organism RTPCR
hey2 WT standard conditions High-pec whole organism RTPCR
WT + MO1-arl4aa standard conditions High-pec whole organism RTPCR
mCherry ci5Tg; um14Tg standard conditions Prim-15 trunk vasculature IFL
ci5Tg; um14Tg + MO1-arl4aa standard conditions Prim-15 trunk vasculature IFL
myb kca3Tg; kca4Tg standard conditions Prim-25 ventral wall of dorsal aorta ISH
kca3Tg; kca4Tg heat shock Prim-25 ventral wall of dorsal aorta ISH
kca3Tg; kca4Tg + MO1-arl4aa standard conditions Prim-25 ventral wall of dorsal aorta ISH
kca3Tg; kca4Tg + MO1-arl4aa heat shock Prim-25 ventral wall of dorsal aorta ISH
kca4Tg heat shock Prim-25 ventral wall of dorsal aorta ISH
kca4Tg + MO1-arl4aa heat shock Prim-25 ventral wall of dorsal aorta ISH
notch1a WT standard conditions High-pec whole organism RTPCR
WT + MO1-arl4aa standard conditions High-pec whole organism RTPCR
notch3 WT standard conditions High-pec whole organism RTPCR
WT + MO1-arl4aa standard conditions High-pec whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-arl4aa standard conditions Prim-15 whole organism gata2a expression decreased amount, abnormal
Prim-15 whole organism gata2b expression decreased amount, abnormal
High-pec whole organism her5 expression decreased amount, abnormal
High-pec whole organism hey2 expression decreased amount, abnormal
High-pec whole organism notch1a expression increased amount, abnormal
High-pec whole organism notch3 expression increased amount, abnormal
ci5Tg; um14Tg + MO1-arl4aa standard conditions Prim-15 trunk vasculature EGFP expression decreased distribution, abnormal
kca3Tg; kca4Tg + MO1-arl4aa standard conditions Prim-25 ventral wall of dorsal aorta myb expression decreased distribution, abnormal
kca3Tg; kca4Tg + MO1-arl4aa heat shock Prim-25 ventral wall of dorsal aorta myb expression increased amount, abnormal
kca4Tg + MO1-arl4aa heat shock Prim-25 ventral wall of dorsal aorta myb expression decreased distribution, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Stem Cell Reports