Figure 1. Analysis of mouse pancreatic CTCs identifies Wnt2 as a candidate CTC gene.
a, Schematic representation of strategy for DGE of RNA isolated from primary mouse pancreatic tumour, metastatic ascites, and CTC-enriched blood. b, Immunofluorescence staining of mouse pancreatic CTC, leukocyte (WBC), and CTC cluster captured on HbCTC-Chip (DAPI, blue; Cytokeratin (CK), red; CD45, green). c, Quantitation of CK+ cells captured from control mice and pancreatic cancer bearing mice (dashed line - threshold of ≥ 6 CK+ cells/100 µL). d, DEGseq M-A plot of RNA sequence reads from mouse (MPANC-9) CTCs, comparing anti-EpCAM-captured versus IgG-coated HbCTC-Chips. (M - log2 fold change, A - averaged log2 reads, red dots – differentially expressed genes) e, RNA-ISH of CTCs and metastatic ascites cells co-expressing CK8+18 (green) and Wnt2 (red) transcripts. White arrowheads highlight Wnt2 signals. f, RNA-ISH of primary pancreatic tumours showing a small cluster of Wnt2 mRNA expressing cells near a pancreatic duct (D) (Wnt2, red dots; hematoxylin, light blue). Hematoxylin and eosin (H&E) stained serial section is shown (right). A second primary tumour imaged under fluorescence demonstrating Wnt2 expression in a subpopulation of tumour cells; high magnification shown (right, Wnt2, red; CK, green; DAPI, blue). (Scale bars=10 µm).