AGS, KATO III, NCI-N87 [N87] and HGC-27 cells were cultured for 24 h in 1% FBS medium and then were trypsinized, resuspended in 1% FBS medium and seeded in 96 multi-well plates. Immunofluorescence experiments were performed as previously described immunolabeling the cells in humidified dark chamber for 2 h with DyLight 554 Phalloidin (Cell Signaling, Beverly, MA, USA). Nuclei were stained with DAPI for 10 min in the dark. After rinsing with PBS, images were acquired with ZOE fluorescent cell imager (Bio-Rad, Milan, Italy)42 (link).
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