Competition binding assay using flow cytometry analysis. Binding of HA-FITC (20 μg/ml) for 1 hr in the presence of modified HA and native HA determined in (A) MDA-MB-231, (B) RKO, (C) PC-3, and (D) LNCaP cancer cell lines. Negative control comprised of incubation of cells with unlabeled HA while positive control cells were incubated only with HA-FITC. The left column indicates fluorescence intensities obtained after competition binding of modified HA polymers during flow analysis, while the right column indicates the normalized fluorescence intensities based on the obtained negative and positive fluorescence intensities. Data are shown as mean ± S.D., * p<0.05, ** p<0.01, *** p<0.001; One-way ANOVA with Tukey multiple comparisons test.