ZFIN ID: ZDB-FIG-150313-8
Casari et al., 2014 - A Smad3 transgenic reporter reveals TGF-beta control of zebrafish spinal cord development. Developmental Biology   396(1):81-93 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 20-25 somites to Long-pec
PHENOTYPE:
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage Range: 20-25 somites to Long-pec

Fig. 6 Smad3/TGFβ signaling is mainly active in post-mitotic cells. (A–B′) Confocal lateral views of immunofluorescence for GFP (green) and phospho-Histone3 (pH3, red) on Tg(12xSBE:EGFP)ia16 embryos injected at 1–2 cell stage either with the control (A), or smad3a-morpholino (B). For each confocal picture (Z-stack), a small brightfield view of a morphant embryo shows which area is displayed (red dashed line). (C–D) Confocal lateral views of immunofluorescence for GFP (green) and pH3 (red) on 2 dpf larva of Tg(12xSBE:EGFP)ia16 line treated with either DMSO (C) or LY364947 (D) at 24hpf. For each confocal picture (Z-stack), a small brightfield view of a 48 hpf larva shows which area is displayed (red dashed line). (C′ and C′′) Confocal zoomed views (single plane) of hindbrain and tail, respectively, of immunofluorescence for GFP (green) and pH3 (red) on DMSO-treated larva of the 12xSBE line. (E–F) Confocal lateral views of the head (E, F) or tail (E′, F′) regions after EdU labeling on 20 hpf embryos treated at 12 hpf either with DMSO (E, E′) or LY364947 (F, F′). For each confocal picture (Z-stack), a small brightfield view of a morphant embryo shows which area is displayed (red dashed line). (G, H) confocal lateral images (Z-stack) of pulse and chase EdU assay on 24 hpf embryos of Tg(12xSBE:EGFP)ia16 line. Embryos have been fixed after a chase of either 2 (G–G′) or 8 (H–H′) hours (hrs) and immunostained for EdU (red) and GFP (green). EdU+ cells fixed after 2 h are roughly in S/G2 phase, while EdU+ cells fixed after 8 h of chase are post-mitotic. (G′, H′) Zoomed view (single plane) of Tg(12xSBE:EGFP)ia16 tail after 2 or 8 h chase. Scale bar is 100 µm in A–B′, E–F′, G, H; 50 µm in C′–C′&prime& 20 µm in G′, H′. Quantification is presented in Supplemental Table S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
EGFP ia16Tg control 20-25 somites tail bud IHC
Long-pec brain IHC
ia16Tg standard conditions Prim-5 caudal fin IHC
Prim-5 head IHC
ia16Tg chemical treatment: LY 364947 20-25 somites Not Detected tail bud IHC
Long-pec Not Detected brain IHC
ia16Tg + MO1-smad3a standard conditions Prim-5 caudal fin IHC
Prim-5 head IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab4-h3 IHC ia16Tg control Long-pec brain
IHC ia16Tg standard conditions Prim-5 caudal fin
IHC Prim-5 head
IHC ia16Tg chemical treatment: LY 364947 Long-pec brain
IHC ia16Tg + MO1-smad3a standard conditions Prim-5 caudal fin
IHC Prim-5 head
Phenotype Details
Fish Conditions Stage Phenotype
ia16Tg chemical treatment: LY 364947 20-25 somites head cell population proliferation increased process quality, abnormal
20-25 somites tail bud cell population proliferation increased process quality, abnormal
Long-pec brain cell population proliferation increased process quality, abnormal
Long-pec caudal fin cell population proliferation increased process quality, abnormal
ia16Tg + MO1-smad3a standard conditions Prim-5 brain cell population proliferation increased process quality, abnormal
Prim-5 caudal fin cell population proliferation increased process quality, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Biology, 396(1), Casari, A., Schiavone, M., Facchinello, N., Vettori, A., Meyer, D., Tiso, N., Moro, E., Argenton, F., A Smad3 transgenic reporter reveals TGF-beta control of zebrafish spinal cord development, 81-93, Copyright (2014) with permission from Elsevier. Full text @ Dev. Biol.