2.4 Luciferase reporter assay
HEK293T cells were treated with KY19382 in 24-well plates for 24 hours. The cells were washed with cold phosphate buffered saline (PBS) and lysed in 55 μl of 1x lysis buffer (Promega, Madison, WI). The cell lysates were centrifuged at 15,920x g at 4°C for 15 minutes.
Thirty microliters of each supernatant was transferred to 96-well plates and 15 μl luciferin was added. The luciferase activity was measured at 485 nm using a FLUOstar OPTIMA luminometer (BMG Labtech, Offenburg, Germany).