Fig. 1.

ZO-1 is required for lumen morphogenesis. (A) Representative images of WT and ZO-1 KD (KD) cysts stained for nuclei (blue, Hoechst 33342), ZO-1 (red), E-cadherin (green) and F-actin (phalloidin, purple) at days 4, 8 and 11 after plating in collagen gels. Yellow arrowheads indicate apical surfaces. Scale bars: 50 µm. (B) Lumen phenotypes at each time point in both growth conditions was classified as filled – i.e. no lumen (green) – multi-lumen (blue) or single lumen (red). Mean±s.e.m. of >100 cysts from at least three independent experiments with three or more samples per condition. *P<0.05 by two-tailed t-test compared to WT. (C) Representative images of WT and KD cysts grown in Matrigel for 8 days; nuclei, blue, Hoechst 33342; F-actin, phalloidin, green. Yellow arrowheads indicate apical surfaces. Scale bar: 10 µm. Data are representative of at least three independent experiments with three or more samples for each condition, all with similar results.