Journal: bioRxiv

Article Title: Feedback regulation between FOXM1 and APC/C Cdh1 determines the changes in cell cycle dynamics during aging

doi: 10.1101/2025.01.26.634919

Figure Lengend Snippet: Mitotic duration becomes steadily correlated with cell cycle length along aging. A. In vitro model of cellular aging. Early passage human dermal fibroblast (HDF) cultures established from skin biopsies of different age donors. B. PIP-FUCCI genetically encoded fluorescence sensor that enables real-time monitoring of distinct cell cycle phases . C. Cell cycle length measured by the time between two consecutive mitoses in neonatal, 10-, 21-, 42- and 87-year-old HDFs expressing the PIP-FUCCI. Mean ± SD are shown as bars. *** p ≤0.001; **** p ≤0.0001 by Mann-Whitney U test, relative to neonatal HDFs. D. Duration of G1-phase, S-phase, G2-phase, and mitosis measured by single live-cell imaging in neonatal to 87-year-old HDFs expressing the PIP-FUCCI biosensor. Mean ± SD are shown as bars. ns nonsignificant; ** p ≤0.01; *** p ≤0.001; **** p ≤0.0001 by Mann-Whitney U test, relative to neonatal HDFs. E. Duration of mitosis measured by the time between nuclear envelope breakdown (NEB) and nuclear envelope reformation (NER) as a function of cell cycle duration in neonatal to 87-year-old HDFs expressing the PIP-FUCCI. The trend lines, Pearson coefficient ( r ) and p -value are shown. n>50 cells, ≥2 independent experiments.

Article Snippet: Human dermal fibroblasts established from skin samples of reported “healthy” Caucasian males with ages ranging from neonatal to octogenarian (neonatal, DFM021711A from ZenBio Biobank; 10-year-old, GM03348; 21-year-old, GM23964; 42-year-old, AG06235; 75-year-old AG11073 and 87-year-old, AG10884; all from Coriell Cell Repository) were cultured in Minimal Essential Medium (MEM) with Earle’s salts and L-glutamine (10-010-CV, Corning, VA, USA), supplemented with 15% Fetal Bovine Serum (FBS) (Gibco, Paisley, UK) and 1x Antibiotic-Antimycotic (Gibco, NY, USA).

Techniques: In Vitro, Fluorescence, Expressing, MANN-WHITNEY, Live Cell Imaging

Journal: PLOS ONE

Article Title: Involvement of an IgE/Mast cell/B cell amplification loop in abdominal aortic aneurysm progression

doi: 10.1371/journal.pone.0295408

Figure Lengend Snippet: (A-D) ROSA MCs were cultured in the presence of conditioned medium from adventitia of control organ donors (NAA) and AAA patients. DARPin ® protein bi53_79 (5 μM) was added (+D) or not (-D) to inhibit IgE binding to MCs’ FcεRI (C-D). (A, C) After 1 hr, degranulation (MFI CD63) was assessed by flow cytometry. The dotted line indicates CD63 MFI in non-stimulated cells. (B, D) After 4 hrs, mRNA IL-4 level was analysed by RT-PCR (2 -ΔΔCt , normalised to RP18S and non-stimulated cells). A,B: Mann-Whitney test; C,D: Wilcoxon matched-pairs signed rank test. (E, F) Concentrations of IL-4, tryptase, and IgE were measured in conditioned medium from AAA adventitia. Pearson correlation analysis (***, p < 0.001).

Article Snippet: Depending on their size, samples were cut in several pieces that were used to prepare conditioned medium, and/or digested for flow cytometry analysis, and/or fixed in 3.7% paraformaldehyde (PFA) for histological studies, and stored in the Inserm human CV biobank (BB-0033-00029), included in the European network BBMRI-ERIC.

Techniques: Cell Culture, Control, Binding Assay, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, MANN-WHITNEY