Figure S1 Structure of the
tsp-15 gene, allele, expression and mutant phenotypes. (A) Mutation, deletion locus and most frequent splicing pattern from the
tsp-15 mutant alleles are shown. The
sv15 mutation is a change in the splice donor site of intron 4, as indicated by the arrow. The regions of deletions in
gk201,
ok854,
ok881, and
tm1666 alleles are indicated by bold lines. Three of these deletions, with the exception of
gk201, result in the lack of most of the second extracellular domain such that these products are no longer functional. In contrast, the
gk201 mutant was indistinguishable from wild type animals (data not shown) despite a 425 bp deletion in the 59 flanking sequence (2703 to 2278) of tsp- 15, indicating that the deletion sequence does not include an essential element for
tsp-15 transcription. (B) The
tsp-15 hypomorph and null mutant,
sv15 has the Dpy and Bli phenotype. Small blisters are indicated by arrows. Homozygotes of the
tsp-15 deletion alleles show identical recessive embryonic lethal phenotypes. Scanning electron microscopy images show the representative
tsp-15 null mutant,
ok881. The
ok881 homozygote is short and fat, showing croissant-like morphology and also has a wrinkled cuticle. Scale bars indicate 50 mm in larva and 10 mm in the embryo. (C) Time lapse images of TSP-15::GFP [51] expression patterns and
tsp-15 null embryos during embryogenesis. Upper panels are confocal images of
tsp-15::gfp expression and Nomarski images of the corresponding embryo. Developmental stages are indicated on the top of the micrographs.
tsp-15::gfp expression was visible in quadrants of cells along the anteroposterior axis. The expression was decreased and body surface expression was visible around the three-fold stage. TSP-15::GFP expression is prominent in lateral hypodermal cells. Lower panels show representative images of
tsp-15(
ok854) null mutant embryos. Each developmental stage is the same as shown in the upper panels. The
tsp-15(
ok854) embryo developed normally, however, at terminal embryogenesis,
tsp-15(
ok854) embryos shrunk and failed to maintain the vermiform shape. Inset represents a different focal plane, which depicts an abnormal body protrusion in
tsp-15 null embryos. Scale bars indicate 10 um.