Figure 3. The effects of neutralizing antibodies to LYVE-1 on LMW-HA-induced migration of LECs.

(A) SVEC4-10 cells were grown on cover slips to 100% confluent monolayers. Sterile pipette tips were used to scratch the confluent monolayer cells to form a 100 μm wound area, and then the cells were cultured for 24 h with or without 3.13 ug/ml LMW-HA. Wells with FGF-2 (20 ng/ml) or CSA (3.13 μg/ml) treatment was included as positive or negative control, respectively. To block LYVE-1/LMW-HA interaction, SVEC4-10 cells were pre-incubated with 10 μg/ml anti-LYVE-1 antibodies or isotype IgG_2A for 2h. Then LMW-HA was added to reach a final concentration of 3.13 μg/ml and incubated for another 24h. Neutralizing anti-LYVE-1 antibodies or isotype IgG only were also included to eliminate the influence of antibodies themselves on the cells. After incubation, the cells were fixed and observed by inverted microscope. All experiments were repeated at least three times and show a representative example. Magnification was ×100. (B) Migration rate (%) was analyzed by Image Pro Plus 6.0 software. ***, p<0.001 compared with the respective control group.