2.10. Adhesion assay
Cells (1 x 104 cells/coverslip) were plated on 13 mm
round glass coverslips and incubated for 1 hour at 37 ÂșC in 5%
CO2 prior to fixation, each sample was studied in
triplicate. The cells were then washed with PBS and fixed for 20 min
with 4% PFA (Sigma, USA) in PBS. The nuclei were stained with DAPI
(1:5,000 dilution, Panreac Applichem A4099, Spain) and all the
coverslips were mounted with Fluoro-Gel medium (Electron Microscopy
Sciences, USA). The images were acquired on an Olympus BX40 microscope
equipped with an Olympus DP73 camera (Spain) at 20X and the total number
of cells attached to the coverslips was quantified.