Fig. 4: DNA-PK phosphorylates cGAS.

a THP-1 cells were mock infected or infected with HSV-1 (MOI = 0.01) for 16 h before co-immunoprecipitation and immunoblotting analysis. b DNA-PK complex and purified cGAS were analyzed by silver staining. c In vitro kinase assay was performed using DNA-PK complex and purified cGAS with or without HT-DNA (1 μg/ml). d 293T cells were transfected with an IFN-β reporter plasmid cocktail with plasmids containing cGAS WT or the mutants and STING. IFN-β activation was determined by luciferase assay. e THP-1 cells stably expressing FLAG-cGAS were mock infected or infected with HSV-1 (MOI = 1) or VSV (MOI = 0.1). WCLs were collected at the indicated hpi and precipitated with anti-FLAG agarose. Precipitated proteins and WCLs were analyzed by immunoblotting with the indicated antibodies. f THP-1 cells stably expressing FLAG-cGAS were mock infected or infected with HSV-1 (MOI = 1) or VSV (MOI = 0.1) and were mock treated or treated with Nu7441 (3 μM). WCLs were collected at 16 hpi and precipitated with anti-FLAG agarose. Precipitated proteins and WCLs were analyzed by immunoblotting with the indicated antibodies. All experiments were done at least twice, and one representative is shown. n = 3 biologically independent samples for d. Data are presented as mean values ± SD. Source data are provided as a Source data file.