Tim knockdown EBs are smaller and of more uniform size.

<p>EBs were grown from control and Tim knockdown ES cell lines Lenti:87-22 and Lenti:89-18 for 6 and 12 days. EBs were fixed, DAPI-stained and imaged with an Olympus F500 confocal microscope. The EB sizes were then estimated by determining the EB surface area from the images using the <u>ImageJ 1.43U software</u> suite. Scattergrams of the resulting data were analyzed using the Kruskal-Wallis test (nonparametric unpaired analysis of variance by ranks; Prizm Software, GraphPad, Inc.) and a significant difference in the median EB size was observed across both groups (P<0.0001). The median EB size is shown in each group by the red bar.