Fig. 4: Mammalian Vps genes essential for STING degradation and termination of type I interferon response.

a, Schematic overview of the screening procedures. b, Screening of mammalian Vps genes required for STING degradation. Sting−/− MEFs reconstituted with mRuby3-STING were treated with siRNA against individual Vps genes, and stimulated with DMXAA for 18 h. Cells were analysed by flow cytometry. MFI of mRuby3 in stimulated cells was divided by MFI of mRuby3 in the corresponding unstimulated cells. The calculated value from cells treated with Vps siRNA was then normalized to that of cells treated with control siRNA. The top 25 genes are highlighted in red. Bright red bars indicate the genes that were also ranked within top 25 in c. c, Screening for mammalian Vps genes required for suppression of STING-dependent type I interferon response. MEFs were treated with siRNA against individual Vps genes, and stimulated with DMXAA for 10 h. Cell supernatants were analysed for type I interferon (IFN). IFN activity from cells treated with Vps siRNA was normalized to that of cells treated with control siRNA. The top 25 genes are highlighted in blue. Bright blue bars indicate the genes that were also ranked within top 25 in b. d, Vps genes ranked within top 25 both in b and c are shown. e, The expression of Cxcl10 in MEFs that were treated with siRNA against the indicated Vps genes, and then stimulated with DMXAA for 12 h. Data are presented as mean ± standard deviation (s.d.). Statistical significances between control siRNA/DMXAA (+) and the indicated siRNAs/DMXAA (+) were determined by performing Student’s unpaired t-test with Bonferroni multiple correction. f, FLAG-STING-reconstituted Sting−/− MEFs were stimulated with DMXAA for 3 h, and lysed. FLAG-STING in the lysates was immunoprecipitated. Co-immunoprecipitated proteins were identified by MS. The ratio of abundance of identified proteins before and after stimulation was then calculated individually. The listed are lysosomal proteins that showed increased abundance after stimulation. Gene Ontology analysis in Uniprot was performed to identify lysosomal proteins. N/A indicates a protein that was not detected without stimulation. The sample size (n) represents the number of the biological replicates (e). Source numerical data are available in source data.