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For microscopic analysis, cells on glass slides were fixed with 4% paraformaldehyde for 24 hours, washed, mounted in VECTASHIELD (Vector Laboratories) and analyzed with a ZEISS Axioskop 40 microscope.
For SEM analysis, cells on substrates were further fixed with 1% osmium tetroxide, dehydrated in a graded series of ethanol/distilled water ranging from 50%to100%0% in steps of 10% for 10 minutes each, and then lyophilized for 1 day.
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For IF analysis, cells grown on monolayer were fixed with 2% buffered paraformaldehyde and permeabilized with 0.2% Triton-X100.
For immunofluorescence analysis, cells grown on glass coverslips were fixed and permeabilized in 3.7% PAF and 0.1%Triton or in methanol for 6 min at -20°C.
For immunofluorescence analysis, cells were grown on glass coverslips inside 6-well plates.
For cell cycle analysis, cells were cultured on substrates with defined rigidity in growth media for five days.
For the colocalization analysis, cells were mounted on slides with culture medium.
For microscopic analysis, cells were plated on non-coated eight-well chamber slides (Nalge Nunc International).
For fluorescence recovery after photobleaching (FRAP) analysis, cells were grown on coverslips in 35-mm dishes.
For cell cycle analysis, cells were seeded on 10-cm diameter plates in RPMI 1640 containing 10% NBCS.
For immunohistochemical analysis, cells were grown on coverslips and thus morphologically very flat and photographed with epifluorescence (not confocal).
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