4.7 Coelenterazine oxidation assay
CTRL and EXP bacteria were collected by centrifugation of 5,000-8,000 r.p.m for 2 minutes and washed twice using PBS. Then, 100 μL PBS culture was added to each well of the 96-well plate, and the mCherry fluorescence level of EXP groups and CTRL groups were adjusted to match each other. Prior to each assay, 10 mM CLZ ethanol solution was diluted in PBS to a final concentration of 250 μM and incubated for 45 minutes to an hour. During the study, 3 μL CLZ diluted solution was added to each well[43]. The luminescence of 490 nm was measured at 21-27℃. CLZ was purchased from Invitrogen (C2944). Thermo Scientific Varioskan LUX was used for quantitating reactions. SkanIt software was used for data management.