Microporous plate photometer

To determine neutrophil ATP levels, a bioluminescence assay kit (S0026, Beyotime, Shanghai, China) was used according to the manufacturer’s protocol. Briefly, the neutrophils were collected and lysed in the lysis buffer, then centrifuged at 12,000 × g for 5 min at 4 °C to collect the supernatant. ATP concentration in the samples was determined by luciferase assay reagent and analyzed using a microporous plate photometer (BioTek Instruments Inc, USA).

ATP was measured using a bioluminescence assay kit (S0026, Beyotime Biotechnology, Shanghai, China). Briefly, the mouse left lung tissues or RAW264.7 cells after modeling from different groups were immediately lysed in the lysis buffer provided with the kit. The supernatant was collected by centrifugation at 12,000 rpm for 15 min at 4°C. The concentration of ATP in the samples was determined by mixing 20 μL supernatant with 100 μL luciferase reagent. A standard curve was prepared using a series of standards at known concentrations. The luminescence of each sample was measured on a microporous plate photometer (BioTek Instruments Inc, USA).