Extended Data Fig. 7: IRF3 docking at STING pS358 facilitates NF-κB activation.
From: STING signals to NF-κB from late endolysosomal compartments using IRF3 as an adaptor
a, Alignment of the C-terminal tail CTT, sequences of STING from different species. b, Wild type THP1 and THP1 STING KO cells were treated with 50 µg/mL cGAMP for the indicated time intervals. Whole-cell lysates were prepared and analyzed by SDS–PAGE followed by immunoblotting with the antibodies shown. c, HaCaT cells treated with Cas9 and scramble gRNA (control), TBK1 gRNAs or IKKƐ gRNAs were stimulated with vehicle or cGAMP for 5 h. Levels of the indicated proteins were determined by immunoblotting. d, Illustration of the design for generation of mice carrying the Sting S357A amino acid substitution (top panel) and mice carrying both S357A and S365A substitutions (bottom panel) by CRISPR/Cas9 microinjection in C57BL/6 J zygotes as described in the Methods section. The bottom part of each panel shows Sanger sequencing chromatograms from genome-edited mice confirming the introduced codon changes. e, RT-qPCR analysis of Il6, and Cxcl10 in total RNA isolated from Wt, StingS357A/S357A, StingS365A/S365A and StingS357A,S365A/S357A,S365A BMMs after stimulation with vehicle, cGAMP (20ug/ml) or diABZ (1uM) for 2 h (n = 3 biological replicates). Results are presented as mean ± SD. P values were calculated using a two-sided, two-way ANOVA with Tukey’s multiple comparisons test. All results presented in this figure are representative from 3 independent experiments with similar results.
