Figure 6
From: CXCR4 expression heterogeneity in neuroblastoma cells due to ligand-independent regulation
Immunofluorescent Staining of CXCR4 and SDF-1 in Neuroblastoma Cells. Cells were seeded onto coverslips and grown to 80% confluency. At desired confluency cells were fixed with 3.7% formaldehyde, permeabilized with 10% NP-40 and blocked with PBG. CXCR4 and SDF-1 immune complexes were generated by incubating cells with 1/1000 dilutions of the mouse anti-CXCR4 monoclonal antibody MAB172 and the goat anti-SDF-1 antibody MAB310 respectively. 1/200 dilutions of FITC-conjugated donkey anti-mouse and TRITC-conjugated donkey anti-goat antibodies were used to localize CXCR4-MAB172 complexes respectively. Immune complexes were visualized using an inverted fluorescent microscope. Figures show superimposed composites of CXCR4 and SDF-1 imaging from the same fields of vision. (A) SK-N-SH cells. (B) SH-SY5Y cells. Result is a representative image taken from ten separate fields of vision.