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ZFIN ID: ZDB-FIG-111201-41
Yang et al., 2011 - Twist controls skeletal development and dorsoventral patterning by regulating runx2 in zebrafish. PLoS One   6(11):e27324 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage Range: 75%-epiboly to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-25 to High-pec

Fig. 4 Morpholino knockdown of twist1a and twist1b induces the appearance of ventralized embryos and increases the expression of runx2b in zebrafish.

A, Antisense morpholinos (MO) for twist1a (twist1a atgMO), twist1b (twist1b atgMO), twist2 (twist2 atgMO) and twist3 (twist3 atgMO), were designed against the 52 UTR and ATG regions, which blocked translation of each transcript. Each atgMO was microinjected into 1-cell to 4-cell embryo and the percentage of each dorsoventral patterning was calculated at 36–48 hpf. B, Representative pictures of induced Class 1–5 ventralized (V1-5/no defect) and dorsalized (D) embryos. V and D phenotype annotations were described in ref. 20 and 21, respectively. C, Table summarizing the ventralized or dorsalized embryo features of zebrafish microinjected with indicated concentration of each atgMO with or without twist1a/1b mRNA or T2 runx2b atgMO. Zebrafish were microinjected with MO-SC, (D) twist1a atgMO or (E) twist1b atgMO with or without (D) twist 1a mRNA or (E) twist 1b mRNA and quantitative RT-PCR for T1 runx2b and T2 runx2b were performed at 8, 14 and 48 hpf (n = 3). Results are shown as the relative expression to β-actin (mean ± SD) and significance was determined by Student′s t-test. (* p<0.05 and ** p<0.01 versus MO-SC; # p<0.05 and ## p<0.01 versus twist1a/1b atgMO).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
runx2b WT control 75%-epiboly whole organism RTPCR
10-13 somites whole organism RTPCR
Long-pec whole organism RTPCR
WT + MO1-twist1b standard conditions 75%-epiboly whole organism RTPCR
10-13 somites whole organism RTPCR
Long-pec whole organism RTPCR
WT + MO2-twist1b standard conditions 75%-epiboly whole organism RTPCR
10-13 somites whole organism RTPCR
Long-pec whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-twist1b standard conditions Prim-25 - High-pec eye absent, abnormal
Prim-25 - High-pec forebrain anterior region non-functional, abnormal
Prim-25 - High-pec notochord absent, abnormal
Prim-25 - High-pec whole organism wholly ventralized, abnormal
WT + MO2-twist1b standard conditions Prim-25 - High-pec eye absent, abnormal
Prim-25 - High-pec forebrain anterior region non-functional, abnormal
Prim-25 - High-pec notochord absent, abnormal
Prim-25 - High-pec whole organism wholly ventralized, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS One for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS One