ZFIN ID: ZDB-FIG-190716-46
Ando et al., 2019 - Cereblon Control of Zebrafish Brain Size by Regulation of Neural Stem Cell Proliferation. iScience   15:95-108 Full text @ iScience
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Antibody:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Prim-5

Fig. 2

Knockdown of crbn or cul4 Impairs Head Development

(A) Schematic structure of CRL4CRBN binding to thalidomide.

(B) Immunoblot analysis (left) and RT-PCR analysis (right) of 24-hpf embryos injected with the indicated MOs against crbn. The indicated antibodies were used to analyze embryos injected with the translation-blocking MO. RT-PCR was performed using the indicated primer set to analyze embryos injected with the splice-blocking MO. The target site of the MO is indicated with red bar in the diagram.

(C and E) One-cell-stage embryos were left uninjected or injected with the indicated MOs against crbn (C) or cul4 (E) with or without corresponding mRNA and then subjected to immunostaining with anti-acetylated α-tubulin antibody at 24 hpf. Fluorescence images (lower panels) and those overlaid with bright-field images (upper panels) are shown.

(D and F) The ratios of head thickness to body length of embryos that were left uninjected or injected with MOs against crbn (D) or cul4 (F) are shown as means ± SD (n = 15 per group). Scale bar, 50 µm. ∗∗∗p < 0.001.

Gene Expression Details No data available
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-tuba IHC WT control Prim-5 forebrain primary neuron
IHC WT + MO1-cul4a control Prim-5 forebrain primary neuron
IHC WT + MO1-cul4a + MO2-cul4b control Prim-5 forebrain primary neuron
IHC WT + MO2-crbn control Prim-5 forebrain primary neuron
IHC WT + MO2-cul4b control Prim-5 forebrain primary neuron
IHC WT + MO3-crbn control Prim-5 forebrain primary neuron
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-cul4a control Prim-5 forebrain primary neuron ab1-tuba labeling decreased amount, abnormal
Prim-5 head decreased size, abnormal
WT + MO1-cul4a + MO2-cul4b control Prim-5 forebrain primary neuron ab1-tuba labeling decreased amount, abnormal
Prim-5 head decreased size, abnormal
WT + MO2-crbn control Prim-5 forebrain primary neuron ab1-tuba labeling decreased amount, abnormal
Prim-5 head decreased size, abnormal
WT + MO2-cul4b control Prim-5 forebrain primary neuron ab1-tuba labeling decreased amount, abnormal
Prim-5 head decreased size, abnormal
WT + MO3-crbn control Prim-5 forebrain primary neuron ab1-tuba labeling decreased amount, abnormal
Prim-5 head decreased size, abnormal
Acknowledgments:
ZFIN wishes to thank the journal iScience for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ iScience