DEV0328545 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Early expression of SoxF and its dependence on wg. SoxF expression is detected by visible (purple, A,B) and fluorescent in situ hybridization (red, C-H) during late L2 (A,B), early (C,F), mid (D,G) and late (E,H) L3 stage either in a wgZ (A,C-E) or rnZ (F-H) background. (C-H) Projections of confocal z-stacks. (B) wgCX3 discs. (A) In late L2 discs, wgZ expression (brown signal) is detected in a small wedge in the anterior ventral wing disc (w). At this stage, SoxF (purple) is expressed in a wider central domain. A similar pattern is detected in the haltere disc (h). (B) A similar stage wgCX3 disc lacks SoxF expression. (C-C′′) SoxF expression is excluded from the wg-expressing cells in the center of the disc − the prospective pouch − before the onset of the expression of wg in the hinge. (D-D′′) SoxF and wg expression remain exclusive once the expression of wg starts in the hinge, a situation that continues during late L3 (E-E′′). During the early (F-F′′), mid (G-G′′) and late (H-H′′) L3, the expression pattern of SoxF is complementary to that of rnZ.

• Supplemental Figure S2 -

  Fig. S2. Mutations in the SoxF gene and the hinge phenotype. (A) Structure of the Drosophila SoxF locus and different SoxF alleles. Pink boxes represent SoxF exons. Two others genes are represented (gray boxes): RpS23 is upstream of SoxF and CG30071 is within its first intron. The SoxF^KG09145 allele carries an insertion of the P(SUPor-P) element in the first intron. This transposon includes two Su(Hw) insulators (I., black boxes), plus the white and the yellow marker genes (red and yellow boxes, respectively). Other alleles were recovered as imprecise excisions of the original P-insertion. The limits of the different deletions generated were mapped approximately by PCR and are indicated by dashed lines, except for the SoxF²⁶ allele, the limits of which were sequenced (brackets). (B,C) In situ hybridization for the SoxF transcript in wing discs. (B) SoxF is expressed in the hinge region of wild-type discs. (C) In SoxF^KG0914 mutant discs, SoxF expression is completely lost. (D) Mutant escapers have a characteristic hinge phenotype with wings held out. (E,F) Higher-magnification views of the hinge of adult wild-type (E) and Sox-mutant (F) wings. The medial and proximal costa (open arrowhead) are fused, the sclerites (asterisk) are disorganized and the alula (black arrowhead) is severely reduced. (G) Detection of the SoxF transcript by RT-PCR. RNA was extracted from SoxF^KG09145 (−) or from wild-type (+) larvae. Primers specific for each of the genes indicated were used to check for the expression of their respective transcripts. Transcripts were detected in (+) and (−) larvae for all genes, except for SoxF in the SoxF^KG09145 (−) extract. These results indicate that the insertion of the P elements results in the specific loss of SoxF gene transcription and, therefore, the SoxF^KG09145 allele can be considered as a transcriptional null.