Journal: bioRxiv
Article Title: Senescent cells secrete chromatin components via senescence-associated extracellular particles
doi: 10.64898/2025.12.12.694055
Figure Lengend Snippet: ( A ) Schematic representation of the experimental approach used to test whether senescent cell–derived small extracellular vesicles and particles (EVPs) can be taken up by proliferating cells. Proliferating IMR90 cells were treated with PKH67, a green fluorescent lipophilic membrane dye, alone or with PKH67-labeled EVPs isolated from senescent cells. ( B ) Representative immunofluorescence images of proliferating cells treated with or without PKH67-labeled sEVP fractions from senescent cells, using different sEVP volumes while keeping the PKH67 lipid dye concentration unchanged. Boxes indicate expanded region and arrows indicate PKH67 puncta in recipient cells. ( C ) Quantification of internalized PKH67-labeled sEVPs by proliferating cells. Data are presented as mean ± SD (n=3 independent experiments). Statistical significance was determined using two-sided one-way ANOVA: *p=0.023, **p=0.0010, ****p<0.0001. ( D ) Schematic representation of the experimental setup: proliferating IMR90 cells were treated with no sEVPs or sEVPs secreted by proliferating cells treated with DMSO, or senescent cells treated with the inhibitor of CCF formation, MDM2 inhibitor RG7388 (MDM2i) or DMSO vehicle. ( E ) Quantification of cGAMP concentration in proliferating cells treated with no sEVPs or sEVPs (40 μL) from proliferating cells treated with DMSO, or senescent cells treated with MDM2i or DMSO vehicle. sEVPs were isolated from equivalent numbers of cells for each condition and resuspended in identical buffer volumes. Data are presented as mean ± SD (n = 3 independent experiments). Statistical significance was determined using two-sided one-way ANOVA: *p=0.011, **p=0.0094.
Article Snippet: The same channel thresholds were maintained for all cells, and the most intense fluorescent overlap was quantified by Imaris as voxels (i.e., overlap between volumetric pixels in either channel).
Techniques: Derivative Assay, Membrane, Labeling, Isolation, Immunofluorescence, Concentration Assay