Updated FTP location. The STING-null RAW264.7 cells expressing TurboID-STING were treated with DMSO or 100 μg/ml DMXAA, and 500 µM biotin was added to the medium. After 1-h incubation, the cells were lysed in 6 M guanidine-HCl containing 100 mM Tris-HCl (pH 8.0) and 2 mM DTT. The lysates were dissolved by heating and sonication, and centrifuged at 20,000 g for 15 min at 4ºC. The supernatants were recovered, followed by reduction in 5 mM DTT at room temperature for 30 min and alkylation in 27.5 mM iodoacetamide at room temperature for 30 min in the dark. Proteins were purified by methanol/chloroform precipitation and solubilized by 0.1% RapiGest SF in 50 mM triethylammonium bicarbonate. After repeated sonication and vortex, the proteins were digested with 20 µg trypsin at 37ºC overnight. The resultant peptide solutions were captured on the anti-biotin antibody-coupled beads in the presence of 1 mg/ml Pefabloc SC during 16 h of incubation at 4ºC. After washing with TBS four times, and with ultrapure water twice, the biotinylated peptides were eluted with 50 µL of 0.1% TFA in 80% acetonitrile three times. The combined eluates were evaporated, and desalted using GL-Tip SDB. The desalted eluates were further evaporated, and redissolved in 0.1% TFA.