Hromosome pairs examined chromosome pairs examinedDNA methylation in B.Biotin N-hydroxysuccinimide ester distachyon chromosomesFig.
Hromosome pairs examined chromosome pairs examinedDNA methylation in B.distachyon chromosomesFig.DNA methylation patterns on chromosomes Bd and Bd of B.distachyon.a FISH with BAC clones ABRH, ABRH, ABRE (red fluorescence).b Distribution of MeC signals around the similar chromosomes.c MeC foci distribution along the longitudinal axes of hugely condensed chromosome pair Bd excised in the metaphase spread shown on a .e MeC signal distribution of Bdhomologues with visible satellite region.g MeC foci arrangement of Bd homologues.Profiles, idiograms and chromosomes Bd d are oriented with their lengthy arm towards the left.Dark green tints on idiograms reflect low methylation level.Methylation profile descriptions as for Fig..DAPI counterstaining, blue fluorescence.Bars mN.Borowska et al.Fig.DNA methylation patterns on mitotic B.distachyon chromosomes after AzaC remedy.a prometaphase chromosomes subjected to .mmolL AzaC.b Methylation pattern of your exact same chromosomes.Positions of centromeres are pointed out by arrows.d FISH with BAC clones ABRH, ABRD and ABRC (red fluorescence) on metaphase chromosomes subjected to .mmolL AzaC.eDistribution of MeC foci around the similar chromosomes.g Prophaseprometaphase chromosomes after .mmolL AzaC therapy.h Methylation pattern with the same chromosomes.c, f, i Superimposed photos of DAPI stained chromosomes and signals of MeC residues.The arrows colour coding redvery higher; yellowhigh and whitelow methylation level.DAPI counterstaining, blue fluorescence.Bars mrDNA site is localised proximally within the lengthy arm of chromosome Bd, though a nucleolar organising area (i.e.containing transcriptionally active S rDNA loci) is located distally inside the quick arm of chromosome Bd (Draper et al.; Garvin et al).Unlike the preceding group, these chromosomes demonstrate extra precise patterns of DNA methylation.Two general types of MeC foci distribution wereapparent for chromosome Bd, based on condensation, one for extremely condensed chromosomes (Fig.a) and yet another 1 for those with clearly visible satellite regions (Fig.e).Each were characterised by the highest levels of DNA methylation in pericentromeric regions, which abruptly decreased towards each chromosome termini.The methylation profile observed in less condensed Bd chromosomesDNA methylation in B.distachyon chromosomesFig.Distinct demethylation of unique B.distachyon chromosomes subjected to .mmolL AzaC.a DAPIstained chromosomes.b Distribution of MeC residues.cSuperimposed photos of DAPI stained chromosomes and mC distribution.Arrow colour coding as for Fig..Bar mshowed considerably reduce methylation at S rDNA sites (Fig.e) than within the very condensed chromosomes (Fig.c).The methylation pattern of chromosome Bd revealed two characteristic peaks of highdensity MeC foci (Fig.g).The initial corresponded using the pericentromeric regions of the chromosome whilst the second was located interstitially on the lengthy arm.Lower in intensity of antiMeC signals in proximal regions of chromosomes Bd was observed.Impact of AzaC on DNA methylation No prominent differences in antiMeC signal distribution had been observed in B.distachyon chromosome complements in the material subjected for the lowest (.mmolL) concentration of AzaC.Immunolocalisation of MeC in metacentric chromosome pairs showed PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/21308498 strong similarity to methylation patterns discovered in chromosomes from the nontreated material (Fig.a).The specific DNA methylation patterns on the smallest submetacentric pairs BdBd have been also retained.In.