Fig. 2
bcd is required for the shape of the egg. (A) dissected ovarioles of bcdCRISPR mothers, showcasing an overall round appearance, along with a single late-staged oocyte (brown arrowhead). (B) egg collection displaying a mixture of wild-type eggs (blue arrowheads) and bcdCRISPR eggs (brown arrowheads). (C) aspect ratio calculated as length/width, as outlined in the figure of dechorionated control embryos (blue bars, n = 26, mean 2.38) and dechorionated bcdCRISPR embryos (brown bars, n = 32, mean 1.83), with measurements following [38]. (D, E) wild-type eggs, presented in lateral (D) and dorsal (E) views to illustrate the morphology of dorsal appendages. (F, G) bcdCRISPR eggs, shown in lateral (F) and dorsal (G) views, revealing shorter but thicker dorsal appendages. (H-I) embryonic stage 15 3D-embryos from a confocal stack using different protein stainings and colors to stain various tissues: b3-Tubulin (green) stains muscles; Runt (yellow) stains neuronal nuclei of the CNS and brain and the hindgut (yellow arrow), a combination of mab BP102 and Vasa (red) stains the ventral chord and germ cell (GC), respectively, of a wild-type embryo in (H) and a bcdCRISPR embryo in (I). Note the irregular duplicated musculature at the anterior and the interrupted CNS in (I). Also observe the asymmetry of the embryo regarding the duplicated posterior end (dashed line). (J) cuticle phenotype from a cross of a bcd RNAi line, bcdHMC04767 x V32 (maternal driver) creating a stronger phenotype than any of the “classical” bcd alleles [2, 36]. (K) bcdCRISPR cuticle exhibiting a rounder shape and a phenotype even stronger than that in (J). (L) extreme, but less frequent bcdCRISPR phenotype. Note the short and round phenotype as a result of the superimposed egg geometry and embryonic patterning phenotype of complete loss of bcd. The red arrow points towards a small circle of denticles infrequently seen in some embryos, see also supplemental Fig. 1G. Dashed lines in (I-L) denote planes of symmetry for duplication of posterior tissues to the anterior side, best seen as duplications of the filzkörper (fk) as a marker for posterior identity. All embryos are oriented anterior to the left and dorsal side up, unless otherwise noted