Author: Ahat, Erpan; Xiang, Yi; Zhang, Xiaoyan; Bekier, Michael E.; Wang, Yanzhuang
Title: GRASP depletion–mediated Golgi destruction decreases cell adhesion and migration via the reduction of a5ß1 integrin Document date: 2019_3_15
ID: rfs7m6or_46
Snippet: HeLa cells were detached with 20 mM EDTA and resuspended in serum-free DMEM. Cells were seeded in Transwell inserts (BD Bio-Coat control 8.0-µm-pore-size inserts, cat. no. 0877121) at a concentration of 5 × 10 4 in 250 µl of serum-free DMEM. In the lower chamber, 750 µl of full growth medium was added as a chemoattractant. After incubation at 37°C for 20 h, cells on the upper side were gently removed by scraping with a cotton swab. The membr.....
Document: HeLa cells were detached with 20 mM EDTA and resuspended in serum-free DMEM. Cells were seeded in Transwell inserts (BD Bio-Coat control 8.0-µm-pore-size inserts, cat. no. 0877121) at a concentration of 5 × 10 4 in 250 µl of serum-free DMEM. In the lower chamber, 750 µl of full growth medium was added as a chemoattractant. After incubation at 37°C for 20 h, cells on the upper side were gently removed by scraping with a cotton swab. The membranes in the inserts were fixed with 4% paraformaldehyde (PFA) for 30 min and stained with 0.5% crystal violet for 1 h, and cells that had migrated to the lower side were visualized under a Leica MZ FLIII stereomicroscope. Images were analyzed with ImageJ software (National Institutes of Health).
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