Extended Data Fig. 1: SARS-CoV-2 induces DNA damage, inflammation and cellular senescence.
A) Immunoblotting of Calu-3, infected (V+ ) or not (V−) with SARS-CoV-2. Calu-3 exposed or not (NT) to HU or IR were used as controls. Infection was monitored by probing for N-protein. B) Quantification of protein levels shown in A; values are relative to V−. C) IF images of HNEpC; nuclei were stained with Hoechst. D) Quantification of the percentage of γH2AX or pRPAS4/8 positive cells shown in C. E) Images of comet assays of Calu-3. Scale bar, 50 μm. F) Quantification of comet tail moment shown in E. Horizontal bars represent the median values ± 95% CI of three independent infections. G) IF images of Calu-3; nuclei were stained with DAPI; arrows point to cGAS+ micronuclei. H) Percentage of cells positive for cGAS dots, micronuclei and cGAS+ micronuclei shown in G. Values are the means ± s.e.m. I) Immunoblots from samples described in Fig. 1A. The experiment was repeated three times with similar results. J) RT–qPCR for pro-inflammatory cytokines expression in Calu-3 and Huh7, 24 h and 48 h after infection, respectively. Values are the means ± s.e.m. of four independent experiments, except for IL6 and IL8 in Huh7 (n = 3) and shown as relative to V−. K) Immunoblotting for cleaved Caspase-3 (cCASP3) from Huh7 lysates. Valproic acid (VPA)-treated HCT116 were used as positive control. Tubulin was used as loading control. The experiment was repeated three times with similar results. L) Images of Huh7 and HNEpC stained for SA-β-gal; HU-treated cells were used as positive control. M) Quantification of positive cells shown in L. N) IF images of HNEpC. O) Percentage of P21- or KI67-positive cells. Scale bar, 10 μm for panels C, G, L, N. Source numerical data and unprocessed blots are available in source data.
