Infinite f2000 pro

The A9 stable cell line in which ELuc was expressed under the control of CAG promoter was seeded in a 96-well clear bottom plate (Nunc, Wiesbaden, Germany) at the density of 1 × 10⁴ cells per well. After 1 day, the medium was replaced with DMEM without phenol red (Gibco-BRL, Grand Island, NY) but supplemented with 10% FBS, 25 mM Hepes/NaOH (pH 7.0, Sigma-Aldrich), and 200 μM D-luciferin (Toyobo), and incubated for 48 h in the presence of various concentrations of SDS (Nacalai Tesque). Bioluminescence was nondestructively measured for 5 s in each well at 37 °C using a microplate-type luminometer (AB-2350 Phelios, ATTO, Tokyo, Japan), and then viability of the same cells was measured using the cell proliferation reagent WST-1 (Takara, Shiga, Japan) according to the manufacturer’s protocol. Absorption was measured at 440 nm using a microplate reader Infinite F2000 PRO (Tecan, Männedorf, Switzerland).

HEK 293 cells expressing the GloSensor (Promega, Madison, WI, USA) and CXCR4 were cultured in a black 96‐well plate (Perkin Elmer, Waltham, MA, USA) for 2–3 days until the cells were confluent. Cells were then incubated with a HBSS/20 mM HEPES/2.5% Luciferin‐EF (Promega) solution for 2 h. Luminescence was determined using a plate reader (infinite F2000PRO; Tecan, Männedorf, Switzerland) until steady state was achieved. Forskolin (1 μM) was added 28 min after the stimulus, and the luminescence was recorded.