Figure 3

HGF promotes invasive motility of single medulloblastoma cells. (A) Single cell motility of DAOY and UW228 cells was measured using live cell imaging (HGF: 20 ng/mL, ARQ197 and PHA-665752 250 nM). Box plots of three independent experiments are shown. (B) Boyden chamber invasion assay under conditions as described in (A). Mean total numbers of cells transmigrated and S.D. of representative triplicate experiment are shown. Statistical analysis: T-test, * = 0.0454, ** = 0.0038. (C) Upper: schematic of microbead invasion assay setup. Lower: microbeads coated with DAOY cells were embedded in collagen and cells were allowed to disseminate for 24 h. Confocal microscopy analysis of LA-EGFP fluorescence 24 h after embedding is shown (left: maximum intensity projection of Z-stacks, right: single cross-section through middle of beads). D) Quantification of mean and range of cell dissemination from microbeads shown in C (triplicate measurements, ten beads quantified per measurement, dot plot with SD). E) High-resolution confocal images of an HGF-induced (20 ng/mL) LA-EGFP expressing DAOY cell migrating in collagen. F-actin distribution is shown as inverted grey scale. Arrow: direction of migration. Note high F-actin content in invasive protrusions at leading edge of the cell.