Figure 2.
Cellular phenotype and Wnt signaling in the CRC subtypes. (a) Heatmap showing association of colon-crypt location (top or base) and Wnt activity in the patient colorectal tumors from the core data sets revealed by applying specific signatures27,28 using the NTP algorithm. In these analyses, statistics include only those samples that were predicted with FDR < 0.2 (see Supplementary Results and Discussion for statistics from all samples including those with FDR > 0.2). (b) TOP-flash assay depicting Wnt activity in CRC cell lines. (c–e) qRT-PCR analysis depicting the average expression of Wnt signaling pathway (c), stem cell (d) and differentiation-specific (e) markers in a set of CRC subtype-representative cancer cell lines (HT29 and LS174T for goblet-like; LS1034, NCI-H508 and SW948 for transit-amplifying; and SW48, HCT8 and SW620 for the stem-like subtype). The qRT-PCR data is plotted relative to the housekeeping gene RPL13A. Error bars represent the s.d. of technical replicates from a representative experiment. (f) Immunostaining analyses for the differentiation markers KRT20 are presented in red, and nuclei are counterstained with DAPI (blue). HCT116 and COLO320 belong to the stem-like, SW1417 and SW948 belong to transit-amplifying, and HT29 and LS174T belong to the goblet-like subtypes. Scale bar represents 15 µm.