Localization and analysis of telomeres in M. incognita chromosomes via FISH

Fluorescent in situ hybridization (FISH) performed on isolated females from infected tomato roots using probe specific to composite telomeric repeats (green) and chromosomes counterstained with DAPI (shown in blue or black and white). A) Localization of telomere repeats on interphase nuclei (a-c) showing the high specificity of FISH staining together with very low background. B) Additional images (a-f) of telomeric chromosomal localization of the Minc composite repeat on metaphases shown on many different chromosomes. DAPI is separately also shown in black and white for better chromosomal end visualization. White arrows represent chromosomes with no visible telomere signal at the end. C) Quantification of chromosome (a) and telomere (b) number manually performed on Figure 4 from main manuscript text showing most of chromosomes have telomere located only at one single end. D) Telomere localization of Minc repeat shown together with enlarged chromosomes and accompanying FISH signals that occupy weaker stained end of chromosomes (regions marked with red arrows). E) Quantification of telomere FISH signals on interphase and prophase nuclei (a-e) showing consistent number of observed telomeres. Spot analysis was done on telomere (green) channel with automatic determination of threshold and spots were counted based on their mutual separation. Panels are shown with merged channels, DAPI shown in black and white, FISH signals and ImageJ quantification respectively. The total number of telomere signals is given in the top right box on last panel for each analyzed nucleus.