The day before the experiments, cells were seeded in six-well plates at concentration of 2.5 × 10 cells per ml.
Before the experiment cells were seeded in RPMI 1640 media without phenol red, and differentiation was activated by overnight incubation with PMA (50 pmol/10 cells).
In brief, 24 hr before the experiment, cells were seeded on a 96-microwell plate (Nunc, Thermo Scientific) in complete growth medium at a density of 2×10 cells/well in triplicate and allowed to adhere overnight at 37°C.
Before the experiment, the seeds were cold-stratified in wet sand in the dark at 4 °C for 3 months and then germinated at a diurnally fluctuating temperature of 25/10 °C (day/night cycle 12 h/12 h with a corresponding light/dark alternation).
One day before the experiments, 30,000 ZMTH3 cells were seeded in black 96-well clear-bottom plates (Corning GmbH, Germany) suitable for fluorescence measurements.
The day before the experiments, 5 × 10 cells were seeded into a 25 cm flask (BD-Falcon Biosciences).
Before the experiments, 1 × 10 cells were seeded into a T25 flask and grown for 5 days.
At 1 day before the experiments, 1 × 10 cells were seeded in 96-well Black/Clear Imaging Plates (BD Biosciences, San Jose, CA, USA) pre-treated with poly--lysine.
Twenty-four hours before the experiment, the cells were seeded in 96-well plates at a concentration of 50 000 cells/well.
U2OS cells expressing functional EGFP-CXCR4 fusion protein were plated in black 96-well plates with optical bottom 18 24 h before the experiment at a seeding density of 8000 cells per well.
