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.