Supplemental Material

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.

This article contains the following supporting material:

• Video 1 - Time-lapse imaging of YFP-ZO-1 during TJ assembly after calcium switch, in control cells.
• Video 3 - Time-lapse imaging of YFP-ZO-1 during TJ assembly after calcium switch, in cells lacking E-cadherin and expressing an E-cadherin-α-catenin fusion construct.
• Video 2 - Time-lapse imaging of YFP-ZO-1 during TJ assembly after calcium switch, in cells lacking E-cadherin.