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. Author manuscript; available in PMC: 2018 Jul 6.
Published in final edited form as: Cell Stem Cell. 2017 Jun 22;21(1):51–64.e6. doi: 10.1016/j.stem.2017.05.020

Figure 2.

Figure 2

BMP2 induces SATB2 and a posterior HOX code in human gut tube spheroids.

(A) Schematic of gut tube spheroid patterning protocol. (B–D) BMP signaling levels as measured by pSMAD1/5/8 (red) staining of spheroids treated with NOGGIN (B), no treatment (C) and BMP2 (D) for 12 hours. (E) pSmad1/5/8 staining of adult mouse colon showing increased BMP signaling at to the top of crypts. (F–H) SATB2 expression in spheroids treated with NOGGIN (F), no treatment (G) and BMP2 (H) for 72 hours. (I) Quantification of the percentage of SATB2+ CDH1+ epithelium following patterning. (J) Principal component analysis of nascent spheroids and spheroids after 3 days of patterning. (K) Gene ontology analysis of differentially expressed genes between BMP vs NOG treated spheroids. (L) Graph of TPM (Transcripts per million) values of spheroids before and after patterning. Samples analyzed were spheroids before patterning (n=2), and NOGGIN, Control and BMP2 treated spheroids 3 days after patterning (n=4 for each group). For quantification in I, 20 organoids from at least 3 experiments were examined. Error bars represent SD. Scale bars = 50 μm. ****p ≤ 0.0001 determined by 2 tailed t-test comparing NOGGIN+Control treated spheroids and BMP2 treated spheroids.