Fig. 1: EHF expression is downregulated in poorly-differentiated colorectal cancers.

A EHF mRNA expression in 1080 cancer cell lines derived from various tumour types obtained from the Broad Institute Cancer Cell Line Encyclopaedia (CCLE) database. B Validation of EHF mRNA expression in 20 CRC cell lines by q-RT-PCR. Data shown are mean ± SEM from a single experiment performed in duplicate. C Haematoxylin and Eosin (H&E) staining of 4 moderately-differentiated (MD) and four poorly-differentiated (PD) CRC cell lines grown as xenografts showing greater presence of glandular structures in MD lines. D mRNA expression of markers of enterocytes, colonic stem cells and colonic lineage determining transcription factors in moderately and poorly-differentiated CRC cell lines. Data shown are the normalised mRNA expression levels from a single RNA analysis per cell line. E Western blots of EHF, VIL1, GPA33 and KRT20 protein in 4 moderately-differentiated and 4 poorly-differentiated CRC cell lines. EHF expression was determined in nuclear lysates, and HDAC-1 expression was assessed as a loading control. Actin was used as a loading control for VIL1, GPA33 and KRT20. F, G Violin plots of the mRNA expression levels of transcription factors implicated in colonic lineage determination in n = 181 moderately-differentiated (MD) and n = 52 poorly-differentiated (PD) primary CRCs. Data were derived from (F) the COAD cohort profiled by the TCGA (The Cancer Genome Atlas) or G the phase III MAX clinical trial cohort [18]. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, Student’s t test.