Fig. 2. MDA-MB-468 and MDA-MB-231 cells express CD44 and exhibit constitutive hyaluronan binding. (A) Cells cultured overnight were detached and stained for flow cytometry. For CD44 expression, cells were incubated either with anti-CD44 monoclonal antibody E1/2 followed by Alexa555 anti-mouse Ig (red graph) or with secondary antibody alone (black graph). For hyaluronan binding, cells were either incubated with FITC-conjugated hyaluronan (FITC-HA, green graph) or medium alone (black graph). (B) MDA-MB-468 cells cultured overnight on glass coverslips were incubated with FITC-hyaluronan for 1 hour at 37°C (green) and then fixed, permeabilised and stained with the anti-CD44 monoclonal antibody E1/2 followed by Alexa555 anti-mouse Ig (red). Nuclei were visualised with TO-PRO-3 (blue). Bar, 50 µm.