Extended Data Fig. 4: SIRT1 enhances innate antiviral response.
a, IFN-β-Luc, PRD I-III-Luc and IFN-α-Luc activity in HEK293T cells transfected with control empty vector (Co.vec), wild-type SIRT1 (wt), or the catalytically inactive SIRT1 mutant (H363Y), followed by infection for 12 h with SeV. b, qPCR analysis of Ifnb1 and Ifna mRNA in HEK293T cells transfected with control empty vector (Co.vec), SIRT1 wt or H363Y and treated with SeV or poly(I:C). c, qPCR analysis of IFNB1 and IFNA mRNA in RAW264.7 cells transfected with control empty vector (Co.vec), SIRT1 wt or H363Y and treated with SeV or poly(I:C) for various time points. d, qPCR analysis of IFNB1 and IFNA1 mRNA in HEK293T cells transfected with control empty vector (Co.vec) or expression plasmid(s) encoding SIRT1 wt or H363Y, IRF3-5D or IRF7 as indicated. e, Bright field microscopy (top) and fluorescence microscopy (bottom) of VSV–GFP in HEK293T cells transfected with indicated control empty vector (Co.vec), SIRT1 wt or H363Y, followed by infection for 12 h with GFP-expressing VSV (MOI, 0.1) (left). Scale bars, 100 μm. The fold change in VSV–GFP intensity was quantified using ImageJ (right). f, qPCR analysis of IFNB1 (far left) and IFNA mRNA (left), VSV RNA (right), and plaque assay of VSV (far right), in HEK293T cells transfected with expression plasmids for SIRT1 wt or H363Y and mock infected (PBS) or infected for 8 h with VSV (MOI, 0.1). Data are representative of three independent experiments (e). n = 3 biological independent samples (a–f). Mean ± s.d., statistical analysis was performed using two-tailed Student’s t-test (a–d, e (right), f).
