Journal: bioRxiv

Article Title: The TIRR–53BP1 axis controls PLK1 spatiotemporal regulation during mitosis

doi: 10.64898/2026.06.29.735294

Figure Lengend Snippet: Representative images of RPE-1 cells (control (CTL)), ΔTIRR, siCTL, siTIRR) exhibiting either ACA-positive (centric) or ACA-negative (acentric) ( a ) lagging chromosomes and ( c ) micronuclei (MN), Quantification of the percentage of cells with ( b ) lagging chromosomes and ( d ) MN (centric/acentric). ( e ) Representative images of DAPI staining showing nuclear morphology of RPE-1 control (CTL), ΔTIRR, siCTL and siTIRR cells, fixed 6 hours post-release from RO-3306-induced arrest at G2/M. ( f ) quantification of nuclear circularity and solidity for the experiment in (e). (g) Bar graphs showing the frequency of mitotic errors (lagging chromosomes, MN, alignment errors) visualized by high resolution live cell imaging of cells progressing through mitosis. (h) Bar graph depicting time from NEBD to anaphase onset determined by live cell imaging of RPE-1 cells as indicated expressing mCherry-H2B and GFP-α-Tubulin, initiated following the release from RO-3306. Data in ( b, d, g ) are mean+/-S.E from three independent experiments ( b n = 150 anaphase cells and d n =1000 cells). Data in ( f ) is represented as violin plot (n =1000 cells) from one representative experiment with three biological replicates. Bar graphs (mean+/-S.D) (h) represent mitotic cells visualized in three independent time-lapse imaging experiments (n > 35 cells each). Unpaired, two-tailed student’s t-test results as indicated. Scale bars as depicted.

Article Snippet: For synchronization in metaphase, cells were at first synchronized in G2/M by treatment with RO-3306. cells were then washed and released into media containing 20μM of proTAME (Medchem Express HY-124955) for 1 hour.

Techniques: Control, Staining, Live Cell Imaging, Expressing, Imaging, Two Tailed Test

Journal: Nature Communications

Article Title: Adhesion-derived condensates control component availability to regulate adhesion dynamics

doi: 10.1038/s41467-026-74001-3

Figure Lengend Snippet: A Representative confocal images of U2OS cells with endogenously tagged TNS1 (mGreenLantern TNS1, mGL-TNS1) seeded for 3 h on different integrin ligands or poly-L-lysine (PLL) as indicated. Images have been deconvolved using Huygens Professional (SVI). Scale bars 20 μm. B Quantification of FA area per cell from n = 56 (FN), 61 (LMN), 97 (COL I), 84 (COL IV), 74 (VTN) and 55 (PLL) cells from two (FN, LMN, COL IV, VTN, PLL) to three (COL I) biological replicates. FAs were quantified as TNS1-positive spots with circularity 0–0.9. C Quantification of the number of circular TNS1 spots (circularity 0.9–1) per cell from n = 56 (FN), 61 (LMN), 97 (COL I), 84 (COL IV), 74 (VTN) and 55 (PLL) cells per condition from two (FN, LMN, COL IV, VTN, PLL) to three (COL I) biological replicates. D Representative confocal images of U2OS cells with mGL-TNS1 seeded either on fibronectin or poly-L-lysine and stained for F-actin and DAPI. Some of the mGL-TNS1 condensates are indicated with arrows. Images have been deconvolved using Huygens Professional (SVI). Scale bar 20 μm. E Maximum intensity projection orthogonal rendering of cells from Fig. 2D. mGL-TNS1 condensates are indicated by arrows. Projection was performed in Huygens Professional (SVI). Scale bar 5 μm. F Representative confocal images of mGL-TNS1 U2OS cells treated with DMSO, blebbistatin, H1152 and Y27632 inhibitors (both ROCK inhibitors). Cells were stained with WGA and DAPI. mGL-TNS1 condensates are indicated with arrows. Scale bars 20 μm. G Quantification of the mGL-TNS1 droplet area and FA area ratio from n = 110 (DMSO), 118 (blebbistatin), 107 (calpeptin), 119 (H1152), 116 (lapatinib), 116 (LY294002), 101 (MG132), 110 (nocodazole), 109 (Ro3306), 112 (saracatinib), 108 (vanadate) and 124 (Y27632) cells from three biological replicates treated with the indicated compounds. Representative images are shown in Fig. 2F and Supplementary Fig. . Statistical analysis was performed using the Kruskal–Wallis test with Dunn’s multiple comparisons test ( B , C , G ). Unless indicated, the statistical significance level is valid for comparison with each of the conditions ( B , C ) or with DMSO ( G ). p < 0.001 (***). Source data, including exact p -values are provided as a file.

Article Snippet: U2OS mGL-TNS1 cells were seeded at FN-coated glass-bottom 24-well plates, left to adhere overnight and treated with either DMSO or indicated compounds as follows: blebbistatin (25 μM, 60 min, StemCell Technologies, 72402); calpeptin (50 μM, 30 min, Selleckchem, S7396); H1152 (5 μM, 120 min, MedChemExpress, HY-15720); lapatinib (0.5 μM, 120 min, MedChemExpress, HY-50898); LY294002 (10 μM, 60 min, Selleckchem, S1105); MG132 (10 μM, 4 h, MedChemExpress, HY-13259); nocodazole (10 μM, 4 h, MedChemExpress, HY-13520); Ro3306 (20 μM, 60 min, MedChemExpress, HY-12529); saracatinib (1 μM, 60 min, Selleckchem, S1006); vanadate (1 mM, 30 min, Sigma, 5086050004); Y27632 (10 μM, 60 min, MedChemExpress, HY-10071).

Techniques: Staining, Comparison