Figure 3 | Scientific Reports

Figure 3

From: Ultra high content analyses of circulating and tumor associated hybrid cells reveal phenotypic heterogeneity

Figure 3

Four rounds of cyCIF to identify and phenotype hybrid cells on patient-matched colorectal cancer (CRC) and pancreatic cancer (PDAC) tumor sections and blood specimens. Each round utilized imaging strands conjugated to Alexa Fluor (AF)546, AF647 and AF750 fluorophores. DAPI nuclear stain was in blue. Round 1: ECAD, EpCAM, panCK, CD45; Round 2: CK19, Ki67, pAKT, CD44; Round 3: CK8, VIM, EGFR; and Round 4: CK7, αSMA, VWF. Hybrid cell in yellow box. Higher magnification, single antigen staining displayed below. Scale bar = 25 µm. αSMA, alpha smooth muscle actin; CK, cytokeratin; CRC, colorectal cancer; ECAD, E-cadherin; EGFR, epidermal growth factor receptor; EpCAM, epithelial cell adhesion molecule; pAKT, phosphorylated protein kinase, strain AK, Thymoma (phosphorylated protein kinase B); panCK, pan-cytokeratin; PDAC, pancreatic ductal adenocarcinoma; VIM, vimentin; VWF, von Willebrand factor.

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