Fig. 1: Senescence is a universal host cell response to viral stress.

a, SA-β-gal, SAHF formation by DAPI and p16^INK4A staining of IMR90 cells 5 days after retroviral infection or mock control. b, GSEA probing senescence-associated genes in WI38 HDFs in VIS and OIS. A positive normalized enrichment score (NES) indicates enrichment compared to proliferating counterparts. Presented NES are significant (false discovery rate (FDR) q < 0.05; see Supplementary Information for individual values). c, Relative expression levels of core senescence and SASP transcripts in cells as in a by RT–qPCR compared to mock control. d, Multiplex bead-based protein analysis in the supernatant of HDF IMR90 (as in a), WI38 and Tig3 cells. MIP-1α is also known as CCL3; MIP-1β is also known as CCL4. e, SA-β-gal staining of IMR90 cells expressing JMJD2C, p53 shRNA (shp53) or control vector. f, Immunoblot analysis of nuclear senescence markers in samples as in d (α-tubulin was used as a loading control). g, Quantification of SA-β-gal positivity in human cell lines (RPE1, A549) infected with AAV, lentivirus, HCoV-NL63 or VSV. h, Quantification of SA-β-gal positivity in human cell lines and primary HNEpC cells infected with VSV-ΔG*-CoV-S, VSV-ΔG*-CoV-2-S or VSV-ΔG*/empty vector (VSV-ΔG*-emp). NIH3T3 cells were used as ACE2-negative, infection-resistant controls. i, SA-β-gal staining of stable hACE2-engineered primary nasal epithelial cells (HNEpC-hACE2) infected with SARS-CoV-2 and treated as indicated at 48 h after infection. GS, GS-441425; UT, untreated. j, Relative expression levels of the indicated transcripts by RT–qPCR of cells as in i, k, GSEA of RNA-seq-based GEP from AECs infected with SARS-CoV-2 compared with uninfected controls (n = 4). Preselected senescence-associated terms with FDR q ≤ 0.01 are shown (see Supplementary Information for individual values). Scale bars, 100 µm (a, e) or 5 µm (SAHF row in a). All quantifications in this figure refer to the mean ± s.d. of n = 3 independent experiments (except k) with individual values as dots in bar plots or representative photomicrographs shown.