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Quantification of EMSA analysis of affinity of Sp-STING1 and Sp-STING3 with the nucleotide second messengers 2'3'-cGAMP, 2'3'-cUA, 3'3'-cGAMP, 3'3'-cGG, and 3'3'-cAA demonstrates that Sp-STING1 preferentially binds 2'3'-linked cyclic dinucleotides and Sp-STING3 preferentially binds 3'3'-linked cyclic dinucleotides. Results are plotted as fraction bound (shifted/total signal) as a function of increasing protein concentration and fit to a single binding isotherm. Data are the mean ± SD of n = 2 or 3 independent experiments.

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