Fig. 3. Mutations in COPA trigger STING-dependent inflammation.

a Representative histograms of flow cytometry analysis for phosphorylated TBK1 (pTBK1, used fluorochrome PE) in monocytes (CD14 + /CD3-) isolated from PBMCs of one COPA syndrome patient (blue)⁶ and two healthy control individuals (HCs) (black, red) before and after treatment with STING inhibitor H-151 (5 µM) for 4 hrs (green line). Dotted line represents isotype control. n = 1. b Quantification of data shown in (a). Bar graph represents fold change in pTBK1 geometric mean fluorescence intensity (MFI) following H-151 treatment relative to untreated control. Data presented as mean of two independent HCs from n = 1 experiments. c Western blot analysis of HEK293T cells following co-overexpression of mCit-STING and COPA mutants E241K and R233H (0.5 µg DNA per construct) 24 hrs after transfection. Representative result of n = 3. d Immunoblot analysis of stably STING-GFP-expressing HEK293T cells 24 hrs after transient transfection with 0.5 µg plasmid DNA encoding COPA WT or mutants. Untransfected cells stimulated with c-di-AM(PS)₂ (20 µM, 2 hrs) are shown as control (last lane). Representative result of n = 3 experiments. e HEK293 cells (express endogenous STING) were transiently transfected with COPA WT or mutants (0.5 µg or 1 µg DNA/well) and harvested for western blot analysis after 24 hrs. As a positive control, cell lysate of THP-1 cells stimulated with HT-DNA (2 µg/ml, 2 hrs) was used. Representative experiment of 3 independent repeats. Source data are provided as a Source Data file for Fig. 3c, d, e.