Cells were fixed with 2% glutaraldehyde and 4% paraformaldehyde (pH 7.3) for 10 min and then washed with 1× PBS. Next, the entire NS membrane with fixed cells was removed from the plastic tube and placed on a glass slide (25 mm by 75 mm) for sample mounting. A droplet of antifading mounting medium (VECTASHIELD) was applied to the cells. Last, the cells on the NS membrane were mounted between the glass slide and a 12-mm-diameter rounded glass coverslip with a thickness of 0.13 to 0.17 mm (Carolina Assistant-Brand). For live-cell imaging, cell samples were detached by trypsinization and washed three times in 1× PBS. The resuspended cells were then placed on an eight-well chamber coverslip. The cell samples were imaged by a spinning disc confocal microscope (Nikon) and were analyzed by ImageJ.

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