Supplementary Figure 5: Effects of UK-5099 in maintaining organoids.

(a) Organoid formation from different regions of small intestine and colon with treatment conditions (n = 6 per condition, p values by t-test with respect to wENR for each location). (b) Loss of passaging efficiency in crypts maintained in wENR compared to UK-5099. When identical cell numbers are passaged normally, wENR treated are more likely to fail to passage. Images are of multiple matrigel/organoid spots of identical size (5uL) on a 6 well plate tiled to make one image (organoids from n = 3 wells were counted and number per well used for quantification, scale bar = 2000 μm). (c) Formation of hyperintense lumens in crypt organoids grown in wENR compared to UK-5099 3 days after changing media to remove CHIR99021 and Valproic acid (n = 69 total organoids from 6 individual wells). (d) Dose curves for individual drugs in supporting organoid formation from crypts (n = 8 wENR, all other conditions n = 6, p value by t-test with respect to wENR). Data are mean ± s.e.m. ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001. All p values were calculated using Student’s t-test.