Fig. 6: The physiological roles of Tsg101 in STING degradation and termination of type I interferon response.
a, MRC-5 cells were treated with siRNAs, and then stimulated with HT-DNA for the indicated times. Cell lysates were analysed by western blot. b, MRC-5 cells were treated with the indicated siRNAs, and then stimulated with HT-DNA. Cells were immunostained with anti-STING (magenta) and anti-pSTING (green) antibodies. c, The fluorescence intensity of STING or pSTING in b was quantified. d, Schematic representation of the experiments with human primary T cells. e, Knockdown efficiency of TSG101 gene in human primary T cells from a representative donor. Data are presented as mean ± s.d. f, The expression of IFIT1 or IFI27 was quantitated with qRT–PCR. Data are presented as mean ± s.d. g, Primary MEFs were treated with siRNAs, and then infected with HSV-1 (MOI 10) for the indicated times. Cell lysates were analysed by western blot. h, Primary MEFs were treated with the indicated siRNAs, and then stimulated with HSV-1 infection (MOI 10) for 8 h. Cells were immunostained with anti-STING (magenta), anti-Lamp1 (green) and anti-ICP4 antibodies. i, The fluorescence intensity of STING in h was quantified. Scale bars, 10 µm (b,h) and 500 nm (magnified images in h). Data are presented in box-and-whisker plots with the minimum, maximum, sample median and first versus third quartiles (c,i). The sample size (n) represents the number of cells (c,i) or the biological replicates (e,f). Source numerical data and unprocessed blots are available in source data.
