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. 2015 Jun 15;142(12):2147–2162. doi: 10.1242/dev.121046

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© 2015. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 4. — Rab8a deficiency-impaired organoid growth is restored by Wnt3a. (A) Crypts (n=100) of each genotype were seeded in triplicate and the number of surviving organoids then counted daily. Growth of 80% of Rab8a^−/− intestinal organoids was arrested within the first 2 days. Rab8a^ΔIEC did not show any significant improvement in organoid survival. Data were collected from three independent experiments. (B,C) After seeding, Rab8^+/+ and Rab8a^−/− intestinal organoids (n=100 for each genotype) were immediately supplemented with Wnt3a or the Gsk3β inhibitor CHIR at the indicated concentrations. Media containing these supplements were replenished daily. Rab8a^−/− intestinal organoids showed similar morphological and proliferative features as Rab8^+/+ organoids upon Wnt3a or CHIR treatment. (D) In the above experiments, surviving organoids were counted daily during 1 week after seeding. Percentages of surviving organoids are plotted. Note that Wnt3a and CHIR significantly increased the number of surviving Rab8a^−/− organoids. Scale bars: 15 μm.