Fig. 2: Real-time measurement of CDN synthesis and determination of CDN levels from cellular extracts. | Nature Communications

Fig. 2: Real-time measurement of CDN synthesis and determination of CDN levels from cellular extracts.

From: A STING-based biosensor affords broad cyclic dinucleotide detection within single living eukaryotic cells

Fig. 2

a DisA enzyme activity assay time course in the presence of increasing concentrations of the 3’3’-c-di-AMP cyclase DisA using BioSTING. b cGAS activity assay in the presence of indicated concentrations of recombinant cGAS with or without Interferon Stimulatory DNA (ISD) using BioSTING. c cGAS activity assay measuring 2’3’-cGAMP production in the presence of fixed cGAS and ISD concentrations with increasing concentrations of the cGAS inhibitor PF-06928215 using BioSTING. d Reaction linear rates calculated from panel c plotted versus PF-06928215 concentration and fit to IC50 curve (Prism 8). e BioSTING FRET response in the presence of methanol extracted cGAMP from HEK293T cells transfected with increasing concentrations of pCDNA3.1-cGAS. f BioSTING FRET response over two-fold serial dilutions of cGAMP methanol extracts from HEK293T cells transfected with 5 µg of pCDNA3.1-cGAS. In panels a and e, individual data points of n = 2 biological replicates are shown. In panels b–d and f, data are presented as mean ±  s.d. of n = 3 (b, f) or n = 8 (c–d) biological replicates.

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