DEV0204083 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 - Adobe PDF

• Supplemental Table S2 - Adobe PDF

• Supplemental Table S3 - Adobe PDF

• Supplemental Table S4 - Adobe PDF

• Supplemental Table S5 - Adobe PDF

• Supplemental Table S6 - Adobe PDF

• Supplemental Table S7 - Adobe PDF

• Supplemental Table S8 - Adobe PDF

• Supplemental Table S9 - Adobe PDF

• Supplemental Figure S1 -

  Fig. S1. Example of the cell counting technique used to quantify cardiomyocytes at 18-somite and 22-somite stages. (A-D) In situ hybridization detects vmhc expression at the 18-somite stage. Dorsal views, anterior towards the top. In cell counting experiments, the number of cardiomyocytes expressing the gene of interest is determined by marking each cell containing NBT/BCIP precipitate. (A,C) Individual cells are easily identified and quantified as the staining is excluded from the nucleus; nuclei are counted if surrounded by precipitate. (B,D) In these examples, the most intensely stained cells are marked with pink dots, the faintest cells are marked with green dots and the intermediate cells are marked with blue dots. All marked cells are included in the total cell count.

• Supplemental Figure S2 - Fig. S2. Inhibition of Hedgehog signaling by cyclopamine treatment is reversible. (A-C) In situ hybridization detects ptc1 expression at the 10-somite stage. Lateral views, anterior towards the top. (A) Robust ptc1 expression is detected at the 10-somite stage in ethanol-treated wild-type embryos. (B) By contrast, ptc1 expression is undetectable in embryos exposed to CyA from the dome stage until the 10-somite stage. (C) Expression of ptc1 is evident in embryos that were exposed to CyA from dome stage until the six-somite stage; following washout at the six-somite stage, embryos were allowed to develop until the 10-somite stage.

• Supplemental Figure S3 -

  Fig. S3. Changes in fli1 expression in embryos with altered Hh signaling. (A-C) In situ hybridization detects expression of the endothelial gene fli1 at the 16-somite stage. Dorsal views, anterior towards the top. Scale bar: 100 µm. (A,B) A subset of anterior endothelial progenitors (arrowheads) appears subtly expanded in CyA-treated embryos. (C) Loss of some anterior endothelial progenitors (arrowheads) is evident in shh-injected embryos.

• Supplemental Figure S4 -

  Fig. S4. MZsmo cells contribute to both chamber populations. Graph depicts number of donor-derived cardiomyocytes detected in individual host embryos. Cases in which donor-derived cells contributed only to the atrium, only to the ventricle, or to both chambers are classified into three separate categories. Bar indicates mean number of myocardial progeny in each category. Both wild-type and MZsmo donor-derived cells contribute to atrial and ventricular host myocardium. Upon contribution to either chamber, similar numbers of donor-derived cardiomyocytes are found regardless of donor genotype. As expected, for both donor genotypes, the hearts with the greatest number of donor-derived cardiomyocytes fall into the category in which cells were found in both chambers, suggesting that more than one donor cell adopted the cardiac progenitor fate.