Live cell time-lapse imaging of YFP-ZO-1 during TJ assembly after calcium switch. A stable cell line of MDCK cells that expresses YFP-ZO-1 was incubated overnight in calcium-free medium. The medium was replaced by pre-warmed F12 medium containing calcium, 20 mM Hepes buffer (pH 7.2) and Oxyrase (to reduce phototoxicity). Imaging was performed on a Zeiss Axiovert S100 TV with a 40x lens (n.a. 0.65). Cells were maintained at 37ºC and images were collected at a rate of 1 frame/min for about 2 hrs. Images were processed in Photoshop to increase contrast and zoomed in to one quarter of the original image.
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