Fig. 3

DLD of gB is critical to mediating cell migration. a, b, d HeLa cells stably expressing gB (d), gBΔR (a, b, d), or gBΔD (d) in 24 well plates when 80-90 % confluent were scratched with a 1000 μl pipette tip. Post scratch, the cells were incubated with medium supplemented with different concentrations of anti-DLD antibodies (a, b), anti-RGD antibodies (d) or pre-immune IgGs (PIG). Wound closure was monitored at 16 h post scratch and imaged with a laser-scanning LSM 510 Carl Zeiss confocal microscope (Magnification, x 20 objective). A representative image of cell migration is provided (a). b, d The open area (scratch) was quantified with TSratch software and the data represented as a histogram. c Data from Transwell migration assays showing alterations on the migration of HeLa cells expressing gBΔR through a permeable membrane in response to incubating them with anti-DLD antibodies is depicted. e MTT cell proliferation assay was performed to monitor the effect of gB on cells. Briefly, target cells were plated to a density of 3,000 cells/well in a 96-well plate in 150 μl volume of growth medium (DMEM supplemented with 10 % FBS). The following day, cells were incubated with DMEM containing 2 % FBS. As a positive control, we supplemented 10 ng/ml of VEGF to one of the wells seeded with untransfected cells. After 72 h, the MTT assay was performed. Each experiment was repeated three times. Columns with different alphabets (panels b, c, d, and e) indicate the values to be statistically significant (p < 0.05) by LSD