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ZFIN ID: ZDB-FIG-130326-16
Lenhart et al., 2013 - Integration of Nodal and BMP Signals in the Heart Requires FoxH1 to Create Left-Right Differences in Cell Migration Rates That Direct Cardiac Asymmetry. PLoS Genetics   9(1):e1003109 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: 20-25 somites
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: 20-25 somites

Fig. 3 Quantitation of Bmp pathway activity by average fluorescence intensity and number of p-Smad1/5/8 positive cells.

Immunofluorescence images for activated Smad1/5/8 (A,B,D,E,G,H,J,K,P–P3) in Tg(myl7:eGFP) embryos (myocardial GFP in B,E,H,K,P–P23) or in Tg(kdrl:egfp) embryos (endocardial GFP in Q–Q3). C,F,I,L: Schematics of B, E, H and I using IMARIS surface tool. GFP signal in green; p-Smad1/5/8 positive cells are labeled according to intensity using the red (low) to yellow (high) spectrum ranging from an intensity value of 35–65, respectively. All images are dorsal views except for optical cross sections generated in IMARIS along the anterior/posterior axis (P2, Q2) or across the myocardium (P3, Q3). A–C: p-Smad1/5/8 fluorescence intensity is higher on the left of the WT cardiac cone (n = 6). D–F: p-Smad1/5/8 fluorescence intensity on both sides of the cone in spaw morphants is similar to that observed in cells on the left in WT (n = 6). G–I: midway mutants exhibit reduced fluorescence intensities and numbers of p-Smad1/5/8 positive cells, while p-Smad levels elsewhere in the embryo are seemingly unaffected (data not shown, n = 3). J–L: bmp4 and spaw double morphants exhibit diminished p-Smad1/5/8 compared with WT (n = 6) but not as severe as observed in midway mutants. M: Comparison of the average fluorescence intensities of p-Smad1/5/8 positive cells on the left and right of the cardiac cone. N: Comparison of the average number of p-Smad1/5/8 positive cells on the left and right of the cardiac cone. O: Comparison of the average number of p-Smad1/5/8 positive cells in the entire cardiac field. P–P3: The myocardium shows little to no overlap of p-Smad1/5/8 positive cells (red) with the green of the myocardium. Q–Q3: Conversely, p-Smad1/5/8 (red) is observed to colocalize with the GFP positive endothelial cells in Tg(kdrl:egfp), indicated with arrowheads in Q2 and Q3, indicating that Bmp signals more strongly to the endocardium. Error bars indicate standard error of the mean. L: Low. H: High. MO: Morpholino. Het: heterozygote.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP foxh1pr1/pr1; twu34Tg standard conditions 20-25 somites heart rudiment myocardium IHC
s843Tg standard conditions 20-25 somites heart rudiment presumptive endocardium IHC
twu34Tg standard conditions 20-25 somites heart rudiment myocardium IHC
twu34Tg + MO3-bmp4 + MO3-spaw standard conditions 20-25 somites heart rudiment myocardium IHC
twu34Tg + MO3-spaw standard conditions 20-25 somites heart rudiment myocardium IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-smad IHC foxh1pr1/pr1; twu34Tg standard conditions 20-25 somites heart rudiment presumptive endocardium
IHC s843Tg standard conditions 20-25 somites heart rudiment presumptive endocardium
IHC twu34Tg standard conditions 20-25 somites heart rudiment presumptive endocardium
IHC twu34Tg + MO3-bmp4 + MO3-spaw standard conditions 20-25 somites heart rudiment presumptive endocardium
IHC twu34Tg + MO3-spaw standard conditions 20-25 somites heart rudiment presumptive endocardium
Phenotype Details
Fish Conditions Stage Phenotype
foxh1pr1/pr1; twu34Tg standard conditions 20-25 somites presumptive endocardium BMP signaling pathway involved in heart development decreased process quality, abnormal
twu34Tg + MO3-bmp4 + MO3-spaw standard conditions 20-25 somites presumptive endocardium BMP signaling pathway involved in heart development decreased process quality, abnormal
twu34Tg + MO3-spaw standard conditions 20-25 somites presumptive endocardium BMP signaling pathway involved in heart development decreased process quality, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.