DEV035451 Supplementary Material

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• Supplemental Figure S1 -

  Fig. S1. Expression of ths and pyr in pyr¹⁸ and ths⁷⁵⁹ homozygous embryos. In situ hybridisation with antisense RNA probes against pyr and ths of embryos from pyr¹⁸ and ths⁷⁵⁹ heterozygous (pyr¹⁸/CyO and ths⁷⁵⁹/CyO) and homozygous (ths⁷⁵⁹/ths⁷⁵⁹ and pyr¹⁸/pyr¹⁸) embryos, respectively. Note that the expression pattern remained unimpaired in the mutants.

• Supplemental Figure S2 -

  Fig. S2. Mesoderm phenotype in ths⁰²⁰²⁶ and pyr⁰²⁹¹⁵ mutants. Embryos were fixed and stained for Eve. (A,B,E,F) Embryonic stage 10; (C,D,G,H) embryonic stage 14. Normal numbers of Eve-positive cells were observed in ths⁰²⁰²⁶ homozygotes (A,C). The number of Eve clusters was reduced, but they were never completely absent, in homozygous pyr⁰²⁹¹⁵ embryos (B,D). In transheterozygous combinations with ths⁷⁵⁹ and pyr¹⁸, ths mutants did not exhibit any defects (E,G), whereas the number of Eve-positive hemisegments was further reduced in pyr transheterozygotes (F,H). For quantification, see Fig. S3.

• Supplemental Figure S3 -

  Fig. S3. Quantitative analysis of complementation of ths and pyr single mutant alleles. (A) Hatching adult homozygous escaper flies from ths and pyr mutant alleles. The number of surviving adult homozygotes (Cy+) was determined and is presented as a percentage of the expected Mendelian number of homozygotes. All crosses were incubated at 25°C. (B) Eve-positive pericardial cell clusters were determined from fixed embryos (stage 10-13) of the phenotypes indicated. Note that the Eve phenotype of pyr²⁹¹⁵ is strongly enhanced in trans to pyr¹⁸.

• Supplemental Figure S4 -

  Fig. S4. Localisation of ths and pyr transcripts as determined by in situ hybridisation. Stage 5 (lateral view) showing expression of ths and pyr in an overlapping domain in the neuroectoderm. Stage 8 (ventral view) showing that during dorsal-lateral migration of the mesoderm, expression of ths remains throughout the entire neuroectoderm (ventral view). Expression of pyr becomes restricted to ventral segmental patches and a strong expression along the dorsal edge of the ectoderm. At stage 9 (lateral view), ths remains expressed in the neuroectoderm. In addition, expression in the delaminating neuroblast becomes obvious. pyr expression is confined to the dorsal edge in the ectoderm. Neuroblast expression begins as neuroblasts delaminate from the neuroectoderm. Stage 10 (lateral view), showing overlapping segmental expression of ths and pyr. The expression domains are strongly overlapping. Stage 10 (ventral view), showing that both ths and pyr are expressed in the neuroblasts.