Figure 4:

Effect of the presence of Neur on cancer attachment, GCX expression, and E-selectin coverage. A and F. Phase contrast images revealing intact and healthy HUVEC monolayers in untreated conditions and after treatment with Neur. B and G. WGA-labeled GCX in UF (B) regions is abundant. Addition of Neur enzyme to the UF environment (G) abolishes WGA-labeled GCX. C and H. Coverage of HUVEC by E-selectin in conditions of isolated UF versus conditions of UF together with Neur enzyme. D and I. Attachment of 4T1 cells to HUVEC in UF region, prior to or after the introduction of 15mU/mL of Neur. E and J. Attachment of MCF7 cells to HUVEC in UF region, prior to or after the introduction of 15mU/mL of Neur. K and L. The quantification of coverage and thickness of GCX labeled by WGA, respectively. M. Coverage of E-selectin in UF with enzyme treatment, compared to UF conditions. N, O, and P. Data quantification of the attachment, clustering and the initial migration of 4T1 and MCF7 breast cancer cells to HUVEC monolayers, respectively. All data are normalized with UF results. Student’s t test was used to compare “UF” vs. “UF + Neur”. Sample sizes are as follows: GCX expression N=3, E-selectin coverage N=6, 4T1 data N=7, MCF7 data N=4. Significance is denoted as *P<0.05, **P<0.01, ***P<0.001, or not significant (ns).