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Fig. 5 | EJNMMI Research

Fig. 5

From: Indium-111-labeled CD166-targeted peptide as a potential nuclear imaging agent for detecting colorectal cancer stem-like cells in a xenograft mouse model

Fig. 5

CD166tp-G18C targeted to CD166-positive CRC cells. a The CD166tp-G18C binding assay in CRC cells was analyzed by flow cytometry. Both CD166+HCT15 and CD166HCT15 cells were treated with CD166tp-G18C-FITC (20 μg/mL) for 1 h. In competitive group, CD166+HCT15 cells were pre-treated with CD166tp-G18C (20 μg/mL) for 1 h and then treated with 20 μg/mL CD166tp-G18C-FITC for 1 h. Data are presented as mean ± SD (n ≥ 3). *P < 0.05, versus (CD166HCT15) CD166tp-G18C-FITC; #P < 0.05, versus (CD166+HCT15) CD166tp-G18C-FITC. b The CD166tp-G18C binding assay in both CD166+HCT15 and CD166HCT15 cells was analyzed by fluorescent microscopy. Both cells were treated with CD166tp-G18C-FITC (20 μg/mL) for 2 h. The total nuclei were stained with 4’, 6-diamino-2-phenylindole (DAPI). Magnification, × 200; scale bar, 100 μm. c CD166tp-G18C and CD166 protein interaction assay. Both CD166+HCT15 and CD166HCT15 cells were treated with CD166tp-G18C-FITC (20 μg/mL) for 1 h. The supernatant of lysates was collected and treated with or without biotinylated-CD166 polyclonal antibody (2 μg/mL) in the presence of streptavidin agarose beads at 4 °C overnight. The fluorescent signaling of immunoprecipitates was analyzed by ELISA. The CD166tp-G18C (20 μg/mL) was pre-treated for competitive inhibition assay. Data are presented as mean ± SD (n ≥ 3). *P < 0.05

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