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Cryo-EM grid preparation and data collection For the cryo-EM grids preparation, 3 l of purified ghrelin-bound complex at 13 mg/ml and GHRP-6-bound ghrelin receptor complex at 8 mg/ml were applied individually onto a glow-discharged holey carbon grid (Quantifoil, Au300 R1.2/1.3) in a Vitrobot chamber (FEI Vitrobot Mark IV)

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The staining solution was prepared according to the kits protocol, and sections were incubated for 60 minutes at 37C in the dark

For example, TNF- and IL-6 reduced the production of NO by inhibiting the phosphorylation of eNOS (80)