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ZFIN ID: ZDB-FIG-190423-5
Li et al., 2018 - Endodermal pouch-expressed dmrt2b is important for pharyngeal cartilage formation.. Biology Open   7(12) Full text @ Biol. Open
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: Prim-25
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Prim-25

Fig. 5 dmrt2b maintains BMP signaling in the pharyngeal region by inhibiting cv2 expression. (A) Expression of BRE-driven EGFP in the pharyngeal region of Tg(BRE:EGFP;sox10:mCherry-CAAX) transgenic embryos injected with 4 ng cMO or dmrt2b MO at 36 hpf. (B,C) p-Smad1/5/8 levels were decreased in the pharyngeal region of dmrt2b mutants (B) and morphants (C) at 36 hpf. Embryos were stained with the indicated antibodies. Nuclei were counterstained with DAPI (blue). (D,E) The expression of cv2 in dmrt2b morphants were examined by in situ hybridization (D) and qRT-PCR (E). All data are presented as the mean of three independent experiments. Error bars represent s.d. Significance was analyzed using unpaired t-tests. **P<0.01. (F) Co-injection of 100 pg cv2 MO with 4 ng dmrt2b MO partially rescued the EGFP expression in Tg(BRE:EGFP;sox10:mCherry-CAAX) transgenic embryos. (G) p-Smad1/5/8 levels were partially rescued in the pharyngeal region of dmrt2b mutants co-injected with 100 pg cv2 MO at 36 hpf. Embryos were stained with the indicated antibodies. Nuclei were counterstained with DAPI (blue). ov, otic vesicle; p-S1/5/8, phosphorylated Smad1/5/8. Scale bars: 50 µm (A–C,F), 20 µm (G).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
bmper TU control Prim-25 pharyngeal pouch ISH
TU + MO3-dmrt2b standard conditions Prim-25 pharyngeal pouch ISH
EGFP pt510Tg ; ioz22Tg control Prim-25 pharyngeal arch IFL
pt510Tg ; ioz22Tg + MO1-bmper + MO3-dmrt2b standard conditions Prim-25 pharyngeal arch IFL
pt510Tg ; ioz22Tg + MO3-dmrt2b standard conditions Prim-25 pharyngeal arch IFL
y1Tg control Prim-25 pharyngeal arch IHC
y1Tg + MO3-dmrt2b standard conditions Prim-25 pharyngeal arch IHC
mCherry pt510Tg ; ioz22Tg control Prim-25 pharyngeal arch IFL
pt510Tg ; ioz22Tg + MO1-bmper + MO3-dmrt2b standard conditions Prim-25 pharyngeal arch IFL
pt510Tg ; ioz22Tg + MO3-dmrt2b standard conditions Prim-25 pharyngeal arch IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-smad IHC TU control Prim-25 pharyngeal arch
IHC dmrt2bioz21/ioz21(TU) standard conditions Prim-25 pharyngeal arch
IHC dmrt2bioz21/ioz21 + MO1-bmper(TU) standard conditions Prim-25 pharyngeal arch
IHC y1Tg control Prim-25 pharyngeal arch
IHC y1Tg + MO3-dmrt2b standard conditions Prim-25 pharyngeal arch
Phenotype Details
Fish Conditions Stage Phenotype
TU + MO3-dmrt2b standard conditions Prim-25 pharyngeal pouch bmper expression increased amount, abnormal
dmrt2bioz21/ioz21(TU) standard conditions Prim-25 pharyngeal arch ab1-smad labeling decreased amount, abnormal
pt510Tg ; ioz22Tg + MO3-dmrt2b standard conditions Prim-25 pharyngeal arch mCherry expression decreased amount, abnormal
Prim-25 pharyngeal arch EGFP expression decreased amount, abnormal
y1Tg + MO3-dmrt2b standard conditions Prim-25 pharyngeal arch EGFP expression decreased amount, abnormal
Prim-25 pharyngeal arch ab1-smad labeling decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Biology Open for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Biol. Open