Figure 4.

PTK7+ Human CoSCs Show Mixed Features of Canonical CoSCs and LRCs

(A) Immunofluorescence staining on formalin-fixed paraffin sections of DAPI (blue), PTK7 (green), and CHGA (red). Scale bar size is indicated.

(B) Experimental scheme. Mice were given three consecutive intraperitoneal injections of 80 mg/kg β-Naphthoflavone (β-NF) within 24 hr. At day 10, animals were killed and small intestinal (SI) crypt cells isolated.

(C) FACS profile of single cell suspension derived from Cyp1a1-H2B-YFP mouse small intestinal crypts. CD24+/lectin-negative cells were divided into YFP-positive label-retaining cells (LRCs) and YFP-negative cells (Non-LRCs).

(D) qRT-PCR expression analysis showing mouse SI label-retaining cells (CD24+/YFP+) are enriched in Chga, Rfx6, Lgr5, and Ptk7 gene expression compared to CD24+/YFP− cells. Shown are two independent experiments (biological replicates) 1 and 2. Each sample was analyzed three times (technical replicate), and error bars indicate SD.

(E) Venn diagram showing genes that overlap between the humanized LGR5 stem cell signature (Hans Clevers Lab) and the humanized label-retaining cell (LRC) signature (Doug Winton Lab).

(F) GSEA analysis was performed using the specific canonical (LGR5) and label-retaining (LRC) cell signatures (see Table S4 and Supplemental Experimental Procedures) for our PTK7 hCoSC microarray datasets (Table S3).

See Figure S4D for an independent experiment performed on a different individual.

(G) qRT-PCR expression analysis showing that PTK7-high human colonic cells are enriched in LRC gene expression when compared with PTK7-low cells. Samples derived from one patient were analyzed three times (technical replicate). Error bars indicate SD (n = 3).

See Figures S4E and S4F for additional supporting data.