DEV022020 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Analysis of the integrated provirus in transgenic birds. (A) The number of proviral insertions in individual G₁ birds was determined by Southern blot analysis of genomic DNA. G₁ birds were scored for GFP expression (+/−) and DNA was digested with EcoRI to generate a unique fragment for each provirus insertion and hybridized with a probe for GFP sequence. Two independent provirus insertions were detected in the six G₁ offspring of cockerel 1-1 identified (birds 1-1:101, 158, 407 and birds 1-1:200, 388, 590), and a single provirus in the one G₁ bird from cockerel 3-11 (bird 3-11:205). (B) The integrity of the transgene for each unique integration event was assayed by digestion with the restriction enzyme StuI that releases an internal proviral fragment containing the CAG-eGFP transgene. Genomic DNA from three G₁ transgenic birds representing the three proviral integrations was digested with StuI and probed with a GFP DNA fragment, to determine the size of the integrated transgene. Bird 3-11:205 contained the full-length transgene. Bird 1-1:158 contained a transgene with a 300 bp deletion, which subsequent sequence analysis revealed to be within the CMV enhancer. Bird 1-1:590 contained a provirus with a 1.3 kb deletion encompassing the TATA box of the transgene. This bird did not express GFP and was not used for subsequent experiments.

• Supplemental Figure S2 -

  Fig. S2. GFP fluorescence in embryos containing two different reporter constructs. GFP fluorescence in G₂ transgenic embryos containing CAG-GFP or CMV-GFP reporter constructs. Images were captured and processed using identical settings.

• Supplemental Figure S3 -

  Fig. S3. The dorsoposterior tail bud contains a mesodermal progenitor cell population in the mouse. (A) The dpTB region from a 10.5-dpc mouse embryo was grafted into a non-fluorescent, 8.5-dpc host and the embryo cultured for 48 hours. The graft generated descendants along body axis with cells remaining in the tail bud. (B) Transverse section of this embryo showing that GFP⁺ cells were present only in the paraxial mesoderm. Blue, nuclear stain. Scale bars: A, 0.5 mm; B, 0.1 mm.