Journal: Nucleic Acids Research

Article Title: Condensins-mediated mitotic folding persists as a structural echo in interphase chromatin

doi: 10.1093/nar/gkag1014

Figure Lengend Snippet: Orthogonal validation of compartment mixing by replicative labeling and microscopy. ( A ) Schematic of the BrdU/EdU pulse-chase labeling strategy. BrdU (1 h) labels early-replicating A-compartment DNA; after a 4 h chase, EdU (20 min) labels late-replicating B-compartment DNA. Cells are arrested in G1 with palbociclib before fixation. ( B ) Representative microscopy images showing DAPI, BrdU, EdU, and merged channels; scale bar: 5 µm. ( C ). Box plots of colocalization metrics (Pearson’s coefficient, overlap coefficient, Manders’ M1, and Manders’ M2) per cell across conditions. Exact P -values from Mann–Whitney U test are shown above brackets. Colocalization analysis: Ncaph–AID n = 546 (DMSO), 954 (Aux); Rad21–AID n = 1, 836 (DMSO), 1, 348 (Aux) cells from two independent experiments.

Article Snippet: Palbociclib (TargetMol, USA) was added to a final concentration of 0.5 μM to prevent G1/S transition for 14–16 h. Cells were then seeded on fibronectin-coated coverslips (40 μg/ml fibronectin, IMTEK, Russia), fixed with 3.7% formaldehyde in PBS for 15 min, permeabilized with 0.5% Triton X-100 in PBS for 20 min, and DNA was denatured with 4N HCl.

Techniques: Biomarker Discovery, Labeling, Microscopy, Pulse Chase, MANN-WHITNEY, Analysis