FIG 1.

Sirt1 interacts with IFI16. (A) HaCaT keratinocytes were transfected with HA-IFI16 for 24 h and then treated with increasing amounts of nicotinamide (NAM; 0, 2, 5, 10 mM) for another 6 h. Afterward, the cells were lysed and subjected to immunoprecipitation (IP) and immunoblot (IB) analysis. (B) PMA-THP1 cells were infected with HSV-1 (MOI = 1) or left uninfected for 12 h and then treated with NAM (5 mM) for 6 h. Afterward, the cells were lysed and subjected to IP and IB analysis. (C and D) HEK293T cells were transfected with various combinations of plasmids as indicated. At 24 h later, IP and IB analysis were performed. The input of IFI16 is shown in panel D, indicated by a star. (E) HEK293T cells were transfected with Myc-Sirt1, together with HA-IFI16 (+) or control vector (−). At 24 h after transfection, the cell lysates were subjected to IP and IB analysis as indicated. (F) HaCaT keratinocytes were transfected with HA-IFI16 and Myc-Sirt1. At 24 h after transfection, the cells were stimulated with HSV-1 or left uninfected for another 8 h. Then, the cells were treated with or without NAM (5 mM) for 6 h. Immunofluorescence was performed using anti-HA (green) and anti-Myc (red). Nuclei were stained with DAPI. Scale bars, 10 μm. (G) PMA-THP1 cells were infected with HSV-1 (MOI = 1) for the indicated periods and then subjected to IP and IB analysis as indicated. (H) Schematic presentation of full-length IFI16. (I) HEK293T cells were transfected with various combinations of plasmids as indicated. At 24 h later, IP and IB were performed. The data are representative of three independent experiments.