Fig. 3. Morphological phenotypes of the different KDs in collagen I gel cyst cultures. (A-H) Samples of (A) control, (B) Stx-3 KD, (C) Cav-1 KD, (D) Gal-3 KD, (E) Anx-13 KD, (F) VIP17 KD, (G) Crb-3 KD and (H) Stx-2 KD cysts were classified according to polarization and lumen formation into three major classes (columns I, II and III). I, cysts with single lumen; II, cysts with multiple lumina; III, cysts with collapsed or no lumen formed (III also includes the cysts with inverted actin meshwork; see the supplementary material, Fig. S2). Cysts were examined after 14 days of culture in collagen I gel with standard MEM supplemented with 5% FCS. Phenotypes were judged on the basis of several different experiments and 100-200 cysts were examined for each KD. Data shown are the mean ± s.d. (n=3). Statistical significance analysis was performed using ANOVA for the three-group comparisons and using Student's t-test for pairwise comparisons. All comparison were either ^*P<0.05 (significant) or ^**P<0.001(very significant). Representative phenotypes for the different KDs are shown in the insets, with actin/TRITC-phalloidin staining in red. Scale bars, 30 µm.