Nature research paper: Cryo-EM structure of an active bacterial TIR–STING filament complex
and trace metals, supplemented with 30 mM nicotinamide to limit TIR toxicity) were grown at 37 °C with 230 r.p.m. shaking. Cultures reaching an optical density at 600 nm > 2.5 were induced with a final IPTG concentration of 500 μM and incubated at 16 °C overnight at 230 r.p.m. Collected bacterial pellets were sonicated in lysis buffer and purified by gravity flow over Ni-NTA resin .
For the second c-di-GMP dataset,was pre-incubated with 1 mM benzamide adenine dinucleotide before rapid mixing with 84 µM c-di-GMP and frozen as above. Semi-automated data collection was performed with SerialEM v3.8.5 and v3.8.6.
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