Figure 3.

In vitro analyses of cGAMP generation upon incubation of cGAS with dsDNA or nucleosomes. (A) Kinetics of cGAMP production induced by 145-nucleosomes and 145-dsDNA. (B,C) Comparison of cGAMP production after incubation of cGAS with different dsDNA and nucleosomes for 3 h. Reaction conditions: dsDNA or nucleosomes (80 ng DNA/μl) were mixed with human cGAS (2 μM) in a Tris–HCl buffer (20 mM, pH 7.6, 60 mM NaCl, 2.5 mM ATP, 2.5 mM GTP, 5 mM MgCl₂). After incubation at 37 °C, aliquots were taken and analysed by UPLC-MS. For all graphs, error bars indicate the mean ± standard deviation of three independent experiments. Statistical significance was determined based on Student’s t-test (ns, p > 0.05; *0.05 > p > 0.01; **0.01 > p > 0.001; ***0.001 > p).