Fig 2: Immunohistochemical analysis of GC tissues and representative images of the immunofluorescence staining of CTCs. (a) Strong expression of EpCAM was detected both on the cell membrane and partially in the cytoplasm of GC cells, and CD147 was identified through intense membranous staining. HCG was positively detected in GC cells with strong cytoplasmic staining or membrane staining. CD45 was negatively detected in GC cells, but was positively expressed on the infiltrating lymphocytes. Red, green and black arrows represent GC cells, smooth muscle cell and infiltrating lymphocytes, respectively. (b) GC CTCs enriched by NanoVelcro system were positive for HCG (coupled with FITC, green) and negative for CD45 (coupled with TRITC red). Blood spiked manually with JEG-3 cells was used as the positive control for HCG. [Color figure can be viewed at wileyonlinelibrary.com]

Fig 3: Characterization of BMSC and osteoblastic differentiation. BMSCs were stained for mesenchymal (CD44 and CD90) and hematopoietic (CD45) cell surface markers (A). Protein is shown with cyan and nucleus with magenta. BMSCs were differentiated to osteoblasts for 12 days and characterized for mRNA levels of Runx2 (B), ALP (C) and OCN (D) (mean ± SD, N = 4–6). BMSCs were used as a control (differentiation day 0). Cyclophilin B was used to normalize the mRNA levels. Statistical significances are shown as *p < 0.05, **p < 0.01 and ***p < 0.001 (One way ANOVA). Cultures were repeated 3 times and representative images are shown. BMSCs and day 6 osteoblasts were stained for osteocalcin (cyan) and nuclei (magenta) (E). Scale bar: 50 µm.