Extended Data Fig. 8: Aged mice show reduced SIRT1 activity and impaired innate antiviral immunity.
a, Cellular SIRT1 activity in PBMCs of non-aged (n = 12, 2–3 months old) and aged (n = 10, 20 months old) mice. b, ELISA of IFN-β in PBMCs from mice as in a infected for 12 h with VSV (MOI, 0.1) (left); Correlation of IFN-β and cellular sirt1 activity in PBMC cells from mice as in a after VSV stimulation (right). c, ELISA of IFN-β in PBMCs from mice as in a infected for 12 h with HSV-1 (MOI, 10) (left); Correlation of IFN-β and cellular sirt1 activity in PBMC cells from mice as in a after HSV-1 stimulation (right). d, IB of TCLs and proteins immunoprecipitated with antibodies to (anti-) acetyl-Lys39 or acetyl-Lys77 of IRF3 (upper) and (anti-) acetyl-Lys45 or acetyl-Lys92 of IRF7 (lower) from PBMCs of non-aged (n = 5) and aged (n = 5) mice, non-infected (−) or infected for 12 h with SeV. e, Immunofluorescence microscopy and DAPI staining of Mouse Pulmonary Fibroblasts (MPF) cells infected for 12 h with SeV. Intensity of intranuclear IRF3 or IRF7 puncta and the percentage of cells showing IRF3 or IRF7 puncta were quantified by ImageJ. Scale bar, 5 μm. f, qPCR analysis of Ifnb1 mRNA in the lungs, spleen and liver of non-aged and aged mice (n = 5 mice per group) given intraperitoneal injection of PBS or VSV (5 × 108 PFU per mouse) for 24 h. g, ELISA of IFN-β in serum from mice as in f. h, qPCR analysis of VSV mRNA in the lungs, spleen and liver of infected mice as in f (left); Plaque assay of VSV in the lungs, spleen and liver of infected mice as in f (right). i, Immunoblot analysis of VSV-G in the liver, lungs and spleen of infected mice as in f. j, Microscopy of hematoxylin-and-eosin (H&E)-stained lung sections from mice as in f. Scale bar, 100 µm. k, Survival rates of non-aged and aged mice (n = 5 mice per group) at various times (horizontal axes) after intraperitoneal infection with VSV (2 × 109 PFU per mouse). l, qPCR analysis of Ifnb1, Cxcl10 and Isg56 mRNA in the brain of non-aged and aged mice (n = 5 mice per group) given intraperitoneal injection of PBS or HSV-1 (5×108 PFU per mouse) for 24 h. m, ELISA of IFN-β in serum from mice as in l. n, qPCR analysis of HSV-1 genomic DNA in brain of mice as in l. o, Plaque assay of HSV-1 in brain of mice as in l. p, Survival rates of non-aged and aged mice (n = 5 mice per group) at various times (horizontal axes) after intraperitoneal infection with HSV-1 (2 × 109 PFU per mouse). Data are representative of at least three independent experiments (d, e, i, j). Mean ± s.d., n = 5 biologically independent animals (g, h, m–o); statistical analysis was performed using two-tailed Student’s t-test (a–c, e–h, l–o) or two-way ANOVA (k, p).
