Fig. 3. cGAS/STING activation by cytoplasmic DNA is also observed in primary human dermal fibroblasts from elderly donors.

a Age group stratification of primary fibroblast bulk RNAseq data (GSE113957; Fleischer et al.²⁴). b Heatmap depicting the expression of cGAS/STING pathway modulators across different age groups. c Enrichment of IFNα and IFNγ responses and TNFα signaling Hallmarks in geriatric fibroblasts. d Interferome analysis (top) showing DEGs that were upregulated in Geriatric fibroblasts, including the proportion of interferon-stimulated genes (ISG, 50.32%); and ISG classification (bottom). e Linear regression analysis of mRNA levels of downstream targets of IRF3 (IFIH1, RTP4) in human fibroblasts during physiological aging (n = 133 samples). f Experimental design: primary culture of normal human dermal fibroblasts (NHDF) from young (28) and old (62 years) donors. Created with BioRender.com. g Senescence-associated ß-galactosidase activity in young and old fibroblasts (n = 151–237 cells). h Representative images showing phospho-Ubiquitin^Ser65 immunostaining (gray) in young and old NHDFs, and corresponding quantification (n = 45–57 cells). i Representative images showing immunostaining of young and old NHDFs for DNA (green) and TOMM20 (magenta, mitochondria), and corresponding quantification (n = 40–55 cells). Scale bars, 25 μm (g, h, i). All data are expressed as the mean ± s.e.m. Dots represent individual samples (e) or cells (h, i). P values were calculated using the Spearman rank correlation (e) or two-tailed Mann–Whitney U-test (h, i). Created with BioRender.com. Source data are provided as a Source Data file.