FIGURE 5.

CD44 and EphA2 colocalize after stimulation of HMVEC with LMW-HA. HLMVEC were cultured on glass slides and incubated with 100 nm LMW-HA for 15 min in serum-free media. Cells were then fixed in 4% paraformaldehyde and double-stained with anti-CD44v10 coupled with Alexa-594 (a and f) and anti-EphA2 coupled with Alexa 488 (b and g) antibodies and mounted using Prolong anti-fade with DAPI (c and h). Images were taken at 20× magnification and merged (d and h) using ImageJ software. Without stimulation, there is minimal CD44v10 (red) colocalization with EphA2 (green). With LMW-HA challenge, there is much greater (yellow) colocalization (Boxed area).