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. 2015 Nov 5;5(6):979–987. doi: 10.1016/j.stemcr.2015.10.003

Figure 3.

Figure 3

PTK7 Specifies Self-Renewing, Multipotent Stem Cells of the Human Colon

(A–D) Immunohistochemistry analysis of (A) PTK7, (B) EPHB2, (C) FABP1, and (D) CK20 expression on serial sections of normal human colonic mucosa. Black arrows indicate specific staining. Scale bar represents 100 μm.

(E) FACS profile of single-cell suspensions from primary human colon stained with antibody to PTK7 (APC-conjugate) (upper). Only epithelial cells were included (EpCAM+) and non-epithelial cells were excluded (CD11−/CD31−/CD45−) from the analysis. Control staining was performed to define PTK7-negative fraction (lower). H, PTK7 high; M, PTK7 medium; L, PTK7 low; N, PTK7 negative.

(F) Representative images of organoid cultures 10 days after seeding equal cell numbers.

(G) Quantification of organoid-forming capacity of colon cells sorted in (E). Shown are data collected from three independent 3D cultures (biological replicates). t test, ∗p value of the average PTK7 high versus average medium lower than 0.05.

(H) qRT-PCR analysis of gene expression on primary human colon epithelial cells purified by FACS according to different PTK7 surface levels. Error bars indicate SD (n = 3 technical replicates, see Figure S3C for an additional supporting experiment).