DEV024349 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Misexpression of Hb generates ectopic RP1/RP4 motoneurons and a thicker motor nerve root. (A) Wild-type embryo stained for HB9 and Fasciclin 3 (FasIII). RP motoneurons express FasIII (chevron) and exit the CNS via the ISNa nerve root (arrowhead). (B) Misexpression of Hb (worniu-gal4 UAS-hb) results in more RP motoneurons and a thicker ISNa fascicle (arrowhead). Dashed vertical lines indicate the edge of the CNS in A and B. (C) Wild-type RP motoneurons project to the indicated lateral muscles (muscle names in white, RP axons labeled in yellow). White arrowheads, nerve roots. Image is one dissected hemi-embryo shown in mirror image, with muscles shown on the right-hand side. (D) Misexpression of Hb (worniu-gal4 UAS-hb) shown with the same orientation and labeling as in C. Nerve roots are thicker (white arrowheads) and RP motoneurons reach their normal muscle field (yellow labels). Scale bars: 3 µm in A,B; 10 µm in C,D.

• Supplemental Figure S2 -

  Fig. S2. Ectopic Isl⁺ HB9⁺ cells that are superficial to RP5 in cas mutant embryos do not express the motoneuron marker Late Bloomer. Islet, green; HB9, magenta; Late Bloomer, red. RP neurons, white arrowheads; ectopic Isl⁺ HB9⁺ cells, yellow arrowhead (1-2 per hemisegment, 90%, n=183). For all panels, a single representative hemisegment of a stage-16 CNS is shown as a maximum intensity projection. Midline, left; anterior, up. A phenotype summary is shown to the right (Late Bloomer, red outlines). Scale bar: 3 µm.

• Supplemental Figure S3 -

  Fig. S3. NB7-3 progeny are unaffected in pdm mutants and after Pdm misexpression. Interneurons of the NB7-3 lineage. Molecular markers shown at top; genotypes shown to left. A schematic of neuronal identity is shown to the right. The first GMC makes the Hb⁺ Kr⁺ EW1 interneuron (and the GW motoneuron, not shown), the second GMC makes the Hb⁻ Kr⁺ EW2 interneuron, and the third GMC makes the Zfh2⁺ EW3 interneuron (Isshiki et al., 2001; Novotny et al., 2002). Pdm is expressed in the EW3 interneuron (Isshiki et al., 2001) and possibly transiently in other neurons in the lineage (Novotny et al., 2002). Anterior, up; midline, left. Scale bar: 3 µm. (A) Wild-type (wt) embryos have EW1-EW3 neurons. (B) pdm mutant embryos have normal EW neuron fates. (C) Pdm misexpression embryos (insc-gal4 UAS-pdm2) have normal EW neuron fates.