Fig. 3: Induction of senescence in primary human astrocytes and co-cultured primary mouse cells with BrdU treatment. | Nature Communications

Fig. 3: Induction of senescence in primary human astrocytes and co-cultured primary mouse cells with BrdU treatment.

From: Systematic profiling reveals distinct senescence signatures and regulators across human brain cell types

Fig. 3: Induction of senescence in primary human astrocytes and co-cultured primary mouse cells with BrdU treatment.

A Representative images of SA β-gal staining in primary human astrocytes following 7-day DMSO or 100 µM BrdU treatment. Quantification of the percentage of SA β-gal positive cells following DMSO (grey) or BrdU (red) treatment (n = 3 replicates). B Confocal images of lamin B1 (green) and Mitotracker Red CMXRos (red) (n = 9 replicates). Scale bars are 33 µm. C Quantification of immunofluorescence staining of nuclear area (square µm), lamin B1 intensity, mitochondrial mass, and mitochondrial membrane potential (intensity) (based on (B), n = 51–115 cells). D Confocal images of γH2AX (green), p21 (red), and Lysotracker Deep Red (white) (n = 9 replicates). Scale bars are 33 µm. E Quantification of immunofluorescence staining of p21 intensity, γH2AX intensity, number of γH2AX foci per cell, and number of lysosomes per cell based on (D) (n = 68–112 cells). F Quantitative PCR of DMSO (grey) and 100 µM BrdU (red) treated primary human astrocytes for CDKN1A, CDKN2A, CDKN2D, and LMNB1 (n = 3 replicates). G Schematic depicting isolation of primary mouse cortical co-cultures with 7-day DMSO or BrdU treatment (Created in BioRender. Plessis-Belair, J. (2025) https://BioRender.com/8ttgr0z). H Representative images of SA β-gal in primary mouse co-cultures with 7-day DMSO control or 100 µM BrdU treatment. Quantification of the percentage of SA β-gal positive cells with DMSO (grey) or BrdU (red) treatment (n = 3 replicates). I Representative confocal images of γH2AX (green), Gfap (red), and Map2 (white) staining. Quantification of γH2AX intensity in Map2+ neurons (top, n = 122 DMSO,199 BrdU cells) and Gfap+ astrocytes (bottom, n = 253 DMSO, 362 BrdU cells). Scale bars are 10 µm. J Representative confocal images of lamin B1 (green) and Gfap (red) staining. Quantification of lamin B1 intensity in Gfap+ astrocytes (n = 314 DMSO, 368 BrdU cells). K Representative confocal images of lamin B1 (green) and Map2 (white) staining. Quantification of lamin B1 intensity in Map2+ neurons (n = 173 DMSO, 184 BrdU cells). Scale bars are 10 µm. Data analyzed by unpaired two-tailed t-test. All graphs show mean with standard deviation (ns, p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001). Source data and exact p-values are provided as a Source Data file.

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