Fig. 1

Affinity purification procedure of telomeric chromatin by a PI polyamide probe (PI-PRICh). a Scheme for telomeric chromatin isolation using a telomere-targeting PI polyamide probe (TH59-DB). The crude chromatin fraction is mixed and incubated with TH59-DB, and the probe-chromatin complexes are isolated by streptavidin affinity purification. The isolated chromatin fractions are analyzed by mass spectrometry for the study of proteins and by next-generation sequencing for the study of ncRNAs. b Chemical structure of TH59-DB. The base recognition profile of TH59-DB is shown in the lower part. c Outline of the plasmid pull-down assay. Linearized plasmids (gray line) with or without the telomeric repeat (thick black line) were mixed with TH59-DB. The mixture was incubated at 37 °C for binding of TH59-DB. Plasmid-TH59-DB hybrids were captured using MyOne C1 streptavidin beads. d Purification of the telomeric repeat-containing plasmid with TH59-DB. (top) Each fraction (Input, Flow-through, and Elution) was analyzed by agarose gel electrophoresis and EtBr staining. The positions of the telomeric repeat-containing plasmid and the empty vector are indicated. (bottom) Bar graph quantifying telomeric DNA capture. Error bars represent standard deviations. e Telomere labeling with TH59-DB in HeLa1.3 cells. Cells were stained with DAPI (first column), TH59-DB (second column) and anti-TRF2 antibody (third column). The merged images are in the fourth column