Figure 3.
STING F269S expression in immune cell lines recapitulates the SAVI patient phenotype. (A and B) Violin plots showing the distribution of the IFN score in THP1 (A) and JTC (B) from qRT-PCR quantification of the median FC of six ISGs 24 h after dox-induced expression of STING WT or STING F269S. Each dot represents one ISG (n = 3 independent experiments; Mann–Whitney test; **, P < 0.01). (C and D) The expression of different inflammatory genes was measured by RT-qPCR 24 h after dox-induced expression of STING WT or STING F269S in THP1 (C) and JTC (D) and expressed as fold versus −dox, normalized to the HPRT1 housekeeping gene (mean ± SD; n = 4 independent experiments; Mann–Whitney test; *, P < 0.05). (E) IP10 levels were measured by ELISA in the supernatant of THP1 24 h after dox induced expression of STING WT or STING F269S (n = 3 independent experiments, ND = not detected). (F) ISGs levels were measured by RT-qPCR 6 h after dox-induced expression of STING F269S in THP1 and expressed as fold versus −dox, normalized to the HPRT1 housekeeping gene (mean ± SD; n = 6 independent experiments; one sample Wilcoxon test; *, P < 0.05). (G) STAT1 phosphorylation was evaluated by western blot in THP1 6 h after dox-induced expression of STING WT or STING F269S. β-ACTIN was used as loading control (n = 2 independent experiments; one representative blot is shown). (H) IFNγ-inducible genes were measured by RT-qPCR 6 h after dox–induced expression of STING F269S in THP1 and expressed as fold versus −dox, normalized to the HPRT1 housekeeping gene (mean ± SD; n = 4/6 independent experiments; one sample Wilcoxon test; *, P < 0.05). (I) Violin plots showing the distribution of the IFN score in THP1 F269S 6 h after treatment with dox ± αIFNAR or αIFNγ. Each dot represents one ISG (n = 4 independent experiments; two-way ANOVA with Dunnett’s multiple comparisons; **, P < 0.01; ns = not significant). (J) IFNγ-inducible genes were measured by RT-qPCR in THP1 F269S 6 h after treatment with dox ± αIFNAR or αIFNγ and expressed as fold versus +dox, normalized to the HPRT1 housekeeping gene (mean ± SD, n = 5 independent experiments; two-way ANOVA with Tukey’s multiple comparisons; *, P < 0.05; **, P < 0.01). Source data are available for this figure: SourceData F3.

STING F269S expression in immune cell lines recapitulates the SAVI patient phenotype. (A and B) Violin plots showing the distribution of the IFN score in THP1 (A) and JTC (B) from qRT-PCR quantification of the median FC of six ISGs 24 h after dox-induced expression of STING WT or STING F269S. Each dot represents one ISG (n = 3 independent experiments; Mann–Whitney test; **, P < 0.01). (C and D) The expression of different inflammatory genes was measured by RT-qPCR 24 h after dox-induced expression of STING WT or STING F269S in THP1 (C) and JTC (D) and expressed as fold versus −dox, normalized to the HPRT1 housekeeping gene (mean ± SD; n = 4 independent experiments; Mann–Whitney test; *, P < 0.05). (E) IP10 levels were measured by ELISA in the supernatant of THP1 24 h after dox induced expression of STING WT or STING F269S (n = 3 independent experiments, ND = not detected). (F) ISGs levels were measured by RT-qPCR 6 h after dox-induced expression of STING F269S in THP1 and expressed as fold versus −dox, normalized to the HPRT1 housekeeping gene (mean ± SD; n = 6 independent experiments; one sample Wilcoxon test; *, P < 0.05). (G) STAT1 phosphorylation was evaluated by western blot in THP1 6 h after dox-induced expression of STING WT or STING F269S. β-ACTIN was used as loading control (n = 2 independent experiments; one representative blot is shown). (H) IFNγ-inducible genes were measured by RT-qPCR 6 h after dox–induced expression of STING F269S in THP1 and expressed as fold versus −dox, normalized to the HPRT1 housekeeping gene (mean ± SD; n = 4/6 independent experiments; one sample Wilcoxon test; *, P < 0.05). (I) Violin plots showing the distribution of the IFN score in THP1 F269S 6 h after treatment with dox ± αIFNAR or αIFNγ. Each dot represents one ISG (n = 4 independent experiments; two-way ANOVA with Dunnett’s multiple comparisons; **, P < 0.01; ns = not significant). (J) IFNγ-inducible genes were measured by RT-qPCR in THP1 F269S 6 h after treatment with dox ± αIFNAR or αIFNγ and expressed as fold versus +dox, normalized to the HPRT1 housekeeping gene (mean ± SD, n = 5 independent experiments; two-way ANOVA with Tukey’s multiple comparisons; *, P < 0.05; **, P < 0.01). Source data are available for this figure: SourceData F3.

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