Ing probe, which was ready from microdissected sex chromatin bodies of WZ females. In females with standard sex chromatin, the Wpainting probe labeled the W Cells 2021, 10, x FOR PEER Evaluation chromosome along its entire length and the telomeric probe hybridized towards the ends of 8 of 22 the WZ BTS 40542 Formula bivalent (Figure 3p,q). On the other hand, in females from two broods with fragmented sex chromatin, only part of the W chromosome was labeled with all the Wpainting probe, and telomeric signals have been detected at each ends of this significant W chromosome as well as painting probe, and telomeric signals were detected at each ends of this substantial W chromoat each ends of its two pairing partners, Z1 and Z2 (Figure 3r,s). These results strongly some as well as at each ends of its two pairing partners, Z1 and Z2 (Figure 3r,s). These recommend that the ancestral W chromosome underwent fusion with an autosome, forming results strongly recommend that the ancestral W chromosome underwent fusion with an aua neoW chromosome. The homologous autosome as a result became the Z2 sex chromosome. tosome, forming a neoW chromosome. The homologous autosome as a result became the Z two No chromosome was differentiated by CGH in male pachytene complements (Figure 3l ). sex chromosome. No chromosome was differentiated by CGH in male pachytene compleThe exact chromosome number could not be identified as a result of an insufficient quantity of ments (Figure 3l ). The exact chromosome number could not be identified as a consequence of an insuitablequality Indole-2-carboxylic acid iGluR mitotic metaphases. sufficient number of suitablequality mitotic metaphases.Figure 3. Sex chromosome systems in Chiasmia clathrata. (a ) Polyploid nuclei stained with orceinFigure 3. Sex chromosome systems in Chiasmia clathrata. (a ) Polyploid nuclei stained with orcein displaying a variable sex chromatin pattern in females from distinct broods, either standard (a) or miniature body (b), whereas it is actually absent in males (c). (d ) Comparative genomeic hybridization (CGH) on pachytene chromosomes revealed a WZ sex chromosome bivalent (arrow and schematic drawing in d) in females with typical sex chromatin (d ) along with a WZ1Z2 trivalent (arrow and schematic drawCells 2021, 10,eight ofshowing a variable sex chromatin pattern in females from various broods, either regular (a) orCells 2021, ten, x FOR PEER Critique (b), whereas it’s absent in males (c). (d ) Comparative genomic hybridization miniature body3.three.(CGH) on pachytene chromosomes revealed a WZ sex chromosome bivalent ((d), arrow and scheme) in females with regular sex chromatin (d ) and a WZ1Z2 trivalent (h, arrow and scheme) in females with scattered sex chromatinWZ bivalent (p,q) and females with theCGH in males (l ).(r,s). Panels (p chromosomes of females with all the (h ); no chromosome was differentiated by WZ1Z2 trivalent Panels (d,h,l)merged photographs of each probes; (e,i,m)female genomic probe (green); (f,j,n)male gemerged photos of both probes; (q,s)DAPI staining (light blue). Within the WZ bivalent, the Wpainting pr nomic probe (red); (g,k,o)DAPI staining (light blue). (p ) Fluorescence in situ hybridization (FISH) labeled theWpaintingchromosome (p, arrow n telomeric probe (green) onZ2 trivalent, less than half from the W with complete W probe (red) and (TTAGG) and scheme). In the WZ1 pachytene chromosomes of females with all the WZ bivalent Wpainting probe, and WZ1 Z2 trivalent (r,s). Panels (p,r)merged chromosome was labeled using the (p,q) and females with all the telomeric signals confirmed the presence of tw images of both probes; (q,s)DAPI staining (ligh.