Fig. 5
Activation of Nrf2 suppresses STING-dependent type IFN response in fibroblasts from SAVI patients. a, b HEK293T were transiently transfected with human WT and SAVI-STING plasmids (V174L, N152S, or V155M) (500 ng/mL). Cells were treated 1 h prior transfection with SFN (10 μM) or 4-octyl-itaconate (4-OI) (200 μM). TMEM173 mRNA levels were assessed by qPCR. Data are the means ± s.e.m. of one experiment performed in quadruplicate. c, d HEK293T were transiently transfected with human WT and SAVI-STING plasmids (V174L, N152S, or V155M) (500 ng/mL), together with an ISRE-Luciferase reporter. Cells were treated 1 h prior transfection of the plasmids with SFN (10 μM) or 4-octyl-itaconate (4-OI) (200 μM). Luciferase activity was assessed 24 h after transfection. Data are the means ± s.e.m. of one experiment performed in duplicate (c) and triplicate (d). e, f Whole-cell lysates from (c) and (d) were analysed by immunoblotting for antiviral signaling with Vinculin (VCL) as loading control. g–i Fibroblasts from SAVI patients were pre-treated with increasing doses of l-sulforaphane (SFN) (10 and 20 μM) for 72 h before getting challenged with cGAMP (4 µg/mL). Whole-cell lysates were analysed for late antiviral signaling events by immunoblotting after 24 h of stimulation. Data are from three individual patients. j Supernatants from (g–i) were collected after 24 h of cGAMP treatment and analysed for type I IFN production. Data are from three individual patients and bars indicated mean and SEM. k–m Immortalized fibroblasts from SAVI patients were pre-treated with 4-OI (200 μM) for 48 h before treating with cGAMP. After 24 h of stimulation cells were collected for western blotting (k–m), and type I IFN analysis by bioassay (n). Data display results from three individual SAVI patients
