Extended Data Fig. 6: AP-1 binds to STING through a dileucine motif.
From: Clathrin-associated AP-1 controls termination of STING signalling
a, HeLa cGAS/STING double KO cells transfected with FLAG-tagged STINGWT, STING1–317 or STINGLI(L364A/I365A) were treated or not with 2.5 µM diABZI for 14 h and analysed by western blot. Vinculin was used as a loading control. b, HeLa cGAS/STING double KO cells transfected with FLAG-tagged STINGWT or STINGL364A were treated or not with 2.5 µM diABZI for 14 h and analysed by western blot. Vinculin was used as a loading control. c, HeLa cGAS/STING double KO cells transfected with FLAG-tagged STINGL363A, STINGL364A or STINGI365A were treated or not with 2.5 µM diABZI for 14 h and analysed by western blot. GAPDH was used as a loading control. d, HEK293T cells transfected with FLAG-tagged STINGWT, STINGL374A or STINGL374F were treated with 2.5 µM diABZI for 2 h, immunoprecipitated with anti-FLAG antibody, and analysed by western blot. e, HeLa cGAS/STING double KO cells transfected with FLAG-tagged STINGWT or STINGL364F were treated or not with 2.5 µM diABZI for 14 h and analysed by western blot. GAPDH was used as a loading control. f, IFN-β luciferase assay in HEK293T cells transfected with plasmids expressing indicated STING constructions and stimulated with diABZI (2.5 µM). g, Glutathione Sepharose pull-down assays of LBD-STINGWT or LBD-STINGELI(E360A/L364A/I365A) by GST-tagged AP-1 ΔμCTD core. One representative of three (a–c,e–f) or two (d,g) independent experiments is shown. Ratios of target proteins versus loading control normalized to the untreated sample of each condition (a–c,e). Mean ± s.d. of three (f) technical replicates. P values based on two-tailed Student’s t-tests (f).
